Preparation of solid cyclodextrin complexes for ophthalmic active pharmaceutical ingredient delivery

ABSTRACT

The present disclosure relates to ophthalmic compositions containing solid complexes of active pharmaceutical ingredient and cyclodextrin, to their method of preparation and their uses. The compositions can include an active agent drug/cyclodextrin complex substantially dissolved in an aqueous eye drop vehicle. The ophthalmic composition is generally in the form of a microsuspension including an active agent complex having a diameter of less than about 100 μm.

CROSS-REFERENCE TO EARLIER APPLICATION

This application claims benefit of U.S. Provisional Patent ApplicationNo. 62/427,737, filed Nov. 29, 2016, incorporated by reference herein inits entirety and relied upon.

FIELD

The present disclosure relates to ophthalmic compositions containingsolid complexes of active pharmaceutical ingredient and cyclodextrin, totheir method of preparation and their uses. The present disclosure alsorelates to bottom-up preparation of novel aqueous eye drop compositionscontaining drug/cyclodextrin nanoparticles.

BACKGROUND

In this specification where a document, act or item of knowledge isreferred to or discussed, this reference or discussion is not anadmission that the document, act or item of knowledge or any combinationthereof was at the priority date, publicly available, known to thepublic, part of common general knowledge, or otherwise constitutes priorart under the applicable statutory provisions; or is known to berelevant to an attempt to solve any problem with which thisspecification is concerned.

According to the National Eye Institute a division of the NationalInstitutes of Health, ocular conditions cause an estimated $139 billioneconomic burden in America alone. This number is not surprisingconsidering that 2.1 million Americans are diagnosed with age-relatedmacular degeneration (AMD), 2.7 million Americans are diagnosed withglaucoma, 7.7 million Americans are diagnosed with diabetic retinopathy,and 24 million Americans are diagnosed with cataracts. Ocular conditionsare not just a problem in the United States. In fact, approximately 285million people worldwide are estimated to be visually impaired.

Most ocular conditions can be treated and/or managed to reduce negativeeffects, including total blindness. To combat this significant problem,the World Health Organization (WHO) approved an action plan with the aimof reducing 25% of the world's avoidable visual impairments by 2019. Inits efforts, the WHO plan to reduce the effects of ocular conditionssuch as diabetic retinopathy, glaucoma, and retinitis pigmentosa, whichaccount for most cases of irreversible blindness worldwide. However,current treatments for ocular conditions are limited by the difficultyin delivering effective doses of drugs to target tissues in the eye.

In current treatments, topical administration of eye drops is thepreferred means of drug administration to the eye due to the convenienceand safety of eye drops in comparison to other routes of ophthalmic drugadministration such as intravitreal injections and implants (LeBourlais, C., Acar, L., Zia, H., Sado, P. A., Needham, T., Leverge, R.,1998. Ophthalmic drug delivery systems—Recent advances. Progress inRetinal and Eye Research 17, 33-58). Drugs are mainly transported bypassive diffusion from the eye surface into the eye and surroundingtissues where, according to Fick's law, the drug is driven into the eyeby the gradient of dissolved drug molecules. The passive drug diffusioninto the eye is hampered by three major obstacles (Gan, L., Wang, J.,Jiang, M., Bartlett, H., Ouyang, D., Eperjesi, F., Liu, J., Gan, Y.,2013. Recent advances in topical ophthalmic drug delivery withlipid-based nanocarriers. Drug Discov. Today 18, 290-297; Loftsson, T.,Sigurdsson, H. H., Konradsdottir, F., Gisladottir, S., Jansook, P.,Stefansson, E., 2008. Topical drug delivery to the posterior segment ofthe eye: anatomical and physiological considerations. Pharmazie 63,171-179; Urtti, A., 2006. Challenges and obstacles of ocularpharmacokinetics and drug delivery. Adv. Drug Del. Rev. 58, 1131-1135).

The first major obstacle is the aqueous drug solubility. In previouslyknown ophthalmic compositions, only dissolved drug molecules canpermeate through biological membranes into the eye. Accordingly,ophthalmic drugs must possess sufficient solubility in the aqueous tearfluid to permeate into the eye.

The second major obstacle is the rapid turnover rate of the tear fluidand the consequent decrease in concentration of dissolved drugmolecules. Following instillation of an eye-drop (25-50 μl) onto thepre-corneal area, the greater part of the drug solution is rapidlydrained from the eye surface and the tear volume returns to the normalresident volume of about 7 μl. Thereafter, the tear volume remainsconstant, but drug concentration decreases due to dilution by tearturnover and corneal and non-corneal drug absorption. The value of thefirst-order rate constant for the drainage of eye drops from the surfacearea is typically about 1.5 min⁻¹ in humans after the initial rapiddrainage. Normal tear turnover is about 1.2 μl/min in humans and thepre-corneal half-life of topically applied drugs is between 1 and 3minutes (Sugrue, M. F., 1989. The pharmacology of antiglaucoma drugs.Pharmacology & Therapeutics 43, 91-138).

The third major obstacle is slow drug permeation through the membranebarrier, i.e. cornea and/or conjunctiva/sclera. The drug molecules mustpartition from the aqueous exterior into the membrane before they canpassively permeate the membrane barrier. The result is that generallyonly few percentages of applied drug dose are delivered into the oculartissues. The major part (50-100%) of the administered dose will beabsorbed from the nasal cavity into the systemic drug circulation whichcan cause various side effects.

The present disclosure seeks to assist with the WHO's plan for reducingavoidable visual impairments by providing an ophthalmic composition anda method of making an ophthalmic composition that overcomes theobstacles of passive drug diffusion in the eye. In these efforts,Applicants provide a method for preparing an ophthalmic composition,which overcomes the major obstacles of passive drug diffusion by (1)increasing the solubility of poorly soluble drugs, (2) increasing theprecorneal half-life of topically applied drugs, and (3) partitioningdrug molecules from the aqueous exterior into the membrane to enablepassive permeation of the membrane barrier. In exemplary embodiments,ophthalmic compositions comprising a combination of such features areprovided.

SUMMARY

Cyclodextrins are well-known to enhance the solubility andbioavailability of hydrophobic compounds. In aqueous solutions,cyclodextrins form inclusion complexes with many active pharmaceuticalingredients. The bottom-up preparation of active pharmaceuticalingredient/cyclodextrin complexes involves suspending an activepharmaceutical ingredient and a cyclodextrin in an aqueous medium andheating the resulting suspension. Upon dissolution of the activepharmaceutical ingredient and the cyclodextrin, complexes of activepharmaceutical ingredient and cyclodextrin are formed. The hot solutionis subsequently cooled to precipitate the solid complexes of activepharmaceutical ingredient and cyclodextrin.

Applicants have surprisingly found that the diameter of the particlesand the viscosity of the composition can be tailored with the heatingand cooling steps and the presence of stabilizing polymers in theaqueous medium. To prevent or substantially inhibit or reduce formationof impurities, such as degradation product derived from the activepharmaceutical ingredient and/or excipients, Applicants have discoveredthat excessive heating of the medium should be avoided and that the hotsolution should be rapidly cooled to room temperature.

Furthermore, the composition of the disclosure exhibits enhancedviscosity which prevents sedimentation of the microparticles duringstorage and also advantageously increases the contact time of theparticles on the surface of the eye thus improving the bioavailabilityof the active pharmaceutical ingredient.

Applicants have discovered an exemplary ophthalmic composition and amethod of making the composition, which overcomes the known majorobstacles of passive drug diffusion.

A first object of the present disclosure is an ophthalmic compositioncomprising, in an ophthalmically acceptable medium, a solid complexcomprising an active pharmaceutical ingredient and a cyclodextrin,wherein the composition comprises less than 2%, in particular less than1%, more particularly less than 0.8%, by weight of impurities based onthe weight of the active pharmaceutical ingredient.

A second object of the present disclosure is an ophthalmic compositioncomprising, in an ophthalmically acceptable medium, a solid complexcomprising an active pharmaceutical ingredient and a cyclodextrin; and apolymer; wherein the viscosity of the composition is from 4 to 14 cP,preferably 5 to 13 cP, more preferably 6 to 12 cP.

A third object of the present disclosure is an ophthalmic compositioncomprising, in an ophthalmically acceptable medium, a solid complexcomprising dexamethasone and γ-cyclodextrin, wherein the compositioncomprises less than 0.5%, in particular less than 0.3%, moreparticularly less than 0.2%, by weight of 16,17-unsaturateddexamethasone or a mixture of enol aldehydes based on the weight of thedexamethasone.

A fourth object of the present disclosure is an ophthalmic compositioncomprising, in an ophthalmically acceptable medium, a solid complexcomprising dexamethasone and γ-cyclodextrin; and a polymer; wherein theviscosity of the composition is from 4 to 14 cP, preferably 5 to 13 cP,more preferably 6 to 12 cP.

The ophthalmic compositions of the disclosure are generally in the formof microsuspensions comprising a solid complex that may exhibit adiameter of less than about 100 μm. The compositions containing naturalα-cyclodextrin, β-cyclodextrin or γ-cyclodextrin and methods provided bythe disclosure provide from about 10-fold to about 100-fold increase inthe concentration of dissolved active pharmaceutical ingredientavailable compared to conventional ophthalmic compositions preparedusing previously-known methods. Moreover, the exemplary methods provideophthalmic composition with decreased concentration of impurities and/orincreased viscosity.

The disclosure also provides methods for preparing ophthalmiccompositions having a high concentration of microparticle activepharmaceutical ingredient/cyclodextrin complex. Moreover, the methods ofthe disclosure provide ophthalmic composition with decreasedconcentration of impurities and/or increased viscosity.

As such, a fifth object of the disclosure is a method of preparing anophthalmic composition, wherein an active pharmaceutical ingredient anda cyclodextrin are suspended in an ophthalmically acceptable medium toform a suspension. The suspension is then heated at a temperature T1lower than 120° C. for a time t until the active pharmaceuticalingredient and the cyclodextrin are substantially dissolved in theophthalmically acceptable medium. The resulting solution is then cooledto a temperature T2 to obtain an ophthalmic composition comprising asolid complex of an active pharmaceutical ingredient and a cyclodextrin.

A sixth object of the disclosure is a method of preparing an ophthalmiccomposition wherein a cyclodextrin is suspended in an ophthalmicallyacceptable medium to form a suspension. The suspension is then heateduntil the cyclodextrin is substantially dissolved in the ophthalmicallyacceptable medium. An active pharmaceutical ingredient is then added insolid form in said solution at a temperature T1 lower than 120° C. andthe mixture is heated at a temperature T1 lower than 120° C. for a timet until the active pharmaceutical ingredient is substantially dissolvedin the ophthalmically acceptable medium. The resulting solution issubsequently cooled to a temperature T2 to obtain an ophthalmiccomposition comprising a solid complex of an active pharmaceuticalingredient and a cyclodextrin.

A seventh object of the disclosure is a method of preparing anophthalmic composition, wherein an active pharmaceutical ingredient issuspended in an ophthalmically acceptable medium to form a suspensionand said suspension is heated until the active pharmaceutical ingredientis substantially dissolved in the ophthalmically acceptable medium.Separately, a cyclodextrin is suspended in an ophthalmically acceptablemedium to form a suspension and said suspension is heated until thecyclodextrin is substantially dissolved in the ophthalmically acceptablemedium. Both compositions are then mixed at a temperature T1 lower than120° C. and the mixture is heated at a temperature T1 lower than 120° C.for a time t. The resulting solution is then cooled to a temperature T2to obtain an ophthalmic composition comprising a solid complex of anactive pharmaceutical ingredient and a cyclodextrin.

An eighth object of the disclosure is a method of preparing anophthalmic composition, wherein dexamethasone and γ-cyclodextrin aresuspended in an ophthalmically acceptable medium to form a suspension.The suspension is then heated at a temperature T1 lower than 120° C. fora time t until the dexamethasone and the γ-cyclodextrin aresubstantially dissolved in the ophthalmically acceptable medium. Theresulting solution is then cooled to a temperature T2 to obtain anophthalmic composition comprising a solid complex of dexamethasone andγ-cyclodextrin.

A ninth object of the disclosure is a method of preparing an ophthalmiccomposition, wherein γ-cyclodextrin is suspended in an ophthalmicallyacceptable medium to form a suspension. The suspension is then heateduntil the γ-cyclodextrin is substantially dissolved in theophthalmically acceptable medium. Dexamethasone is then added in solidform in said solution at a temperature T1 lower than 120° C. and themixture is heated at a temperature T1 lower than 120° C. for a time tuntil the dexamethasone is substantially dissolved in the ophthalmicallyacceptable medium. The resulting solution is subsequently cooled to atemperature T2 to obtain an ophthalmic composition comprising a solidcomplex of dexamethasone and γ-cyclodextrin.

An tenth object of the disclosure is a method of preparing an ophthalmiccomposition, wherein dexamethasone is suspended in an ophthalmicallyacceptable medium to form a suspension and said suspension is heateduntil the dexamethasone is substantially dissolved in the ophthalmicallyacceptable medium. Separately, γ-cyclodextrin is suspended in anophthalmically acceptable medium to form a suspension and saidsuspension is heated until the γ-cyclodextrin is substantially dissolvedin the ophthalmically acceptable medium. Both compositions are thenmixed at a temperature T1 lower than 120° C. and the mixture is heatedat a temperature T1 lower than 120° C. for a time t. The resultingsolution is then cooled to a temperature T2 to obtain an ophthalmiccomposition comprising a solid complex of dexamethasone andγ-cyclodextrin.

An eleventh object of the disclosure is an ophthalmic compositionobtainable by a method according to the disclosure.

A twelfth object of the disclosure is an ophthalmic compositionaccording to the disclosure or prepared according to the method of thedisclosure for use in the treatment of an ocular condition, inparticular an anterior ocular condition or a posterior ocular condition,more particularly uveitis, macular edema, macular degeneration, retinaldetachment, ocular tumors, fungal or viral infections, multifocalchoroiditis, diabetic retinopathy, proliferative vitreoretinopathy(PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome,histoplasmosis, uveal diffusion, and vascular occlusion.

A thirteenth object of the disclosure is an ophthalmic compositionaccording to the disclosure or prepared according to the method of thedisclosure for use in the treatment of macular edema, wherein thecomposition is topically administered to the eye in an amount of 1 dropof composition three times per day.

A fourteenth object of the disclosure is a use of an ophthalmiccomposition according to the disclosure or prepared according to themethod of the disclosure as an eye drop solution.

In exemplary embodiments, an ophthalmic composition comprises an activeagent drug/cyclodextrin complex dissolved in an aqueous eye dropvehicle. The ophthalmic composition is generally in the form of amicrosuspension comprising an active agent complex having a diameter ofless than about 100 μm. The compositions and methods provided by theexemplary embodiments provide about 10 to 100 fold increase in theconcentration of dissolved active agent (i.e. drug) available inconventional ophthalmic compositions prepared using previously-knownmethods. Moreover, the exemplary methods provide ophthalmic compositionwith decreased concentration of degradation product.

The term “active agent,” as used herein, can also be referred to, forexample, as a pharmaceutical ingredient, an active pharmaceuticalingredient, an ophthalmic active pharmaceutical ingredient or a drug (orvariations thereof). And, as used herein these terms (and variationsthereof) can be considered equivalent and interchangeable. Exemplaryembodiments provide methods for preparing ophthalmic compositions havinga high concentration of microparticle active agent/cyclodextrin complex,and that does not generate or produce a by-product and/or degradationproduct for at least about 90 days when stored at room temperature.

According to one method, an active agent (or drug or other equivalentterm) and at least one cyclodextrin are suspended in an aqueous eye dropvehicle to provide a suspension having a milky appearance. Thesuspension is heated for a sufficient time at a sufficient temperatureuntil the drug and cyclodextrin are dissolved in the aqueous eye dropsolution, and no or substantially no degradation and/or by-product isformed. Once the drug and the cyclodextrin are dissolved, the milkysuspension turns into a substantially clear solution. The resultingsolution is cooled at a rate sufficient to produce a microsuspensioncomprising a microparticle drug/cyclodextrin complex.

In another method, an active agent, at least one cyclodextrin, and atleast one polymer are suspended in an aqueous eye drop vehicle toprovide a suspension having a milky appearance. The suspension is heatedfor a sufficient time at a sufficient temperature until the drug,cyclodextrin, and the polymer are dissolved or substantially dissolvedin the aqueous eye drop solution, and no degradation or substantially nodegradation and/or by-product is formed. Once the drug, thecyclodextrin, and the polymer are dissolved the milky suspension turnsinto a substantially clear solution. The resulting solution is cooled ata rate sufficient to produce a microsuspension comprising amicroparticle drug/cyclodextrin/polymer complex.

In a further method, at least one cyclodextrin is suspended in anaqueous eye drop vehicle to provide a suspension having a milkyappearance. The suspension is heated for a sufficient time at asufficient temperature until the cyclodextrin is dissolved (orsubstantially dissolved) in the aqueous eye drop solution, and nodegradation or substantially no degradation and/or by-product is formed.Once the cyclodextrin is dissolved the milky suspension turns into asubstantially clear solution. An active agent is added to the heatedaqueous suspension, while stirring the solution, until the drug isdissolved or substantially dissolved in the solution. The resultingsolution is cooled at a rate sufficient to produce a microsuspensioncomprising a microparticle drug/cyclodextrin complex.

In yet another method, cyclodextrin and at least one polymer aresuspended in an aqueous eye drop vehicle to provide a suspension havinga milky appearance. The suspension is heated for a sufficient time at asufficient temperature until the cyclodextrin and the polymer aredissolved or substantially dissolved in the aqueous eye drop solution,and no degradation or substantially no degradation and/or by-product isformed. Once the cyclodextrin and the polymer are dissolved, the milkysuspension turns into a substantially clear solution. An active agent isadded to the heated aqueous suspension, while stirring the solution,until the drug is dissolved in the solution. The resulting solution iscooled at a rate sufficient to produce a microsuspension comprising amicroparticle drug/cyclodextrin/polymer complex.

In still another method, at least one cyclodextrin is suspended in wateror an aqueous eye drop vehicle to provide a suspension having a milkyappearance. Separately the drug is suspended in water or an eye dropvehicle free of cyclodextrin to provide a suspension having a milkyappearance. The two suspensions are sterilized by, for example, heatingin an autoclave at 121° C. for 20 minutes. Then the two suspensions orhot solutions are allowed to cool to about 95° C. before mixing to forma substantially clear solution, and no degradation or substantially nodegradation and/or by-product is formed. The resulting solution iscooled at a rate sufficient to produce a microsuspension comprising amicroparticle drug/cyclodextrin complex.

In a non-limiting embodiment, at least one cyclodextrin is suspended inwater or an aqueous eye drop vehicle to provide a suspension having amilky appearance. Separately, the active pharmaceutical ingredient issuspended in water or an eye drop vehicle free of cyclodextrin toprovide a suspension having a milky appearance. The two suspensions areheated or sterilized by, for example, heating in an autoclave at 121° C.for 20 minutes. Then the two suspensions or hot solutions are mixedtogether and the temperature is adjusted to about 90° C. to about 95° C.to form a substantially clear solution, and no degradation orsubstantially no degradation and/or by-product is formed. The resultingsolution is cooled at a rate sufficient to produce a microsuspensioncomprising a microparticle drug/cyclodextrin complex.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features of this disclosure will now be described withreference to the drawings of certain embodiments which are intended toillustrate and not to limit the disclosure.

FIG. 1 depicts the structures of the natural α-cyclodextrin,β-cyclodextrin, and γ-cyclodextrin.

FIG. 2 depicts the effect of self-aggregation of γ-cyclodextrin onactive pharmaceutical ingredient (dexamethasone) solubilization.Phase-solubility profiles of dexamethasone and the naturalγ-cyclodextrin (γCD) in aqueous eye drops. The solid curve is thesolubility of the active pharmaceutical ingredient (•), the broken curveis the solubility of γCD (∘), and the straight line is the theoreticalamount of dissolved γCD in the aqueous eye drop medium. Excess amount ofdexamethasone was added to a solution containing 0 to 20% (w/v) γCD inan ophthalmically acceptable medium containing benzalkonium chloride(0.02% w/v), sodium edetate (0.1% w/v) and sufficient sodium chloride toobtain isotonic solution.

Thus, the amount of drug dissolved in solution is constant at γCDconcentrations above 3% (w/v). The amount of γCD dissolved in solutionincreases slower than the amount of γCD that is added to the media, thechange only becomes linear after 10% (w/v). This shows that at γCDconcentrations between about 3-10% (w/v), all γCD added to the mediaforms solid complexes with the drug and precipitates. At γCDconcentrations above 10% (w/v) the amount of dissolved γCD shows again alinear increase.

DETAILED DESCRIPTION

Further aspects, features and advantages of the exemplary embodimentswill become apparent from the detailed description which follows.

The patents, published applications and scientific literature referredto herein establish the knowledge of those with skill in the art and arehereby incorporated by reference in their entireties to the same extentas if each was specifically and individually indicated to beincorporated by reference. Any conflict between any reference citedherein and the specific teachings of this specification shall beresolved in favor of the latter. Likewise, any conflict between anart-understood definition of a word or phrase and a definition of theword or phrase as specifically taught in this specification shall beresolved in favor of the latter.

As used herein, whether in a transitional phrase or in the body of aclaim, the terms “comprise(s)” and “comprising” are to be interpreted ashaving an open-ended meaning. That is, the terms are to be interpretedsynonymously with the phrases “having at least” or “including at least”.When used in the context of a method, the term “comprising” means thatthe method includes at least the recited steps, but may includeadditional steps. When used in the context of a composition, the term“comprising” means that the composition includes at least the recitedfeatures or components, but may also include additional features orcomponents.

The terms “consists essentially of” or “consisting essentially of” havea partially closed meaning, that is, they do not permit inclusion ofsteps or features or components which would substantially change theessential characteristics of a method or composition; for example, stepsor features or components which would significantly interfere with thedesired properties of the compounds or compositions described herein,i.e., the method or composition is limited to the specified steps ormaterials and those which do not materially affect the basic and novelcharacteristics of the method or composition.

The terms “consists of” and “consists” are closed terminology and allowonly for the inclusion of the recited steps or features or components.

As used herein, the singular forms “a,” “an” and “the” specifically alsoencompass the plural forms of the terms to which they refer, unless thecontent clearly dictates otherwise.

The term “about” is used herein to mean approximately, in the region of,roughly, or around. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth. In general, the term“about” or “approximately” is used herein to modify a numerical valueabove and below the stated value by a variance of 20%.

The term “dissolved” or “substantially dissolved” is used herein to meanthe solubilization of a solid in a solution. It can be considered that asolid is “dissolved” or “substantially dissolved” in a solution when theresulting solution is clear or substantially clear.

The term “clear” is used herein to mean a translucent or asubtranslucent solution. Thus, a “clear” solution has a turbiditymeasured according to ISO standards of 100 Nephelometric Turbidity Units(NTUs), preferably 50 NTUs.

The term “substantially clear” is used herein to mean a translucent or asubtranslucent solution. Thus, a “substantially clear” solution has aturbidity measured according to ISO standards of 00 NephelometricTurbidity Units (NTUs).

As used herein, the term “cloudy” or “substantially cloudy” or refers toa solution having a turbidity measured according to ISO standards ofgreater than 100 NTUs.

As used herein, the term “milky” or “substantially milky” refers to asolution having a turbidity measured according to ISO standards ofgreater than 100 NTUs, preferably greater than 200 NTUs.

As used herein, the recitation of a numerical range for a variable isintended to convey that the variable can be equal to any values withinthat range. Thus, for a variable which is inherently discrete, thevariable can be equal to any integer value of the numerical range,including the end-points of the range. Similarly, for a variable whichis inherently continuous, the variable can be equal to any real value ofthe numerical range, including the end-points of the range. As anexample, a variable which is described as having values between 0 and 2,can be 0, 1 or 2 for variables which are inherently discrete, and can be0.0, 0.1, 0.01, 0.001, or any other real value for variables which areinherently continuous.

In the specification and claims, the singular forms include pluralreferents unless the context clearly dictates otherwise. As used herein,unless specifically indicated otherwise, the word “or” is used in the“inclusive” sense of “and/or” and not the “exclusive” sense of“either/or.”

Technical and scientific terms used herein have the meaning commonlyunderstood by one of skill in the art to which the present descriptionpertains, unless otherwise defined. Reference is made herein to variousmethodologies and materials known to those of skill in the art. Standardreference works setting forth the general principles of pharmacology andpharmaceutics include Goodman and Gilman's The Pharmacological Basis ofTherapeutics, 10^(th) Ed., McGraw Hill Companies Inc., New York (2001)and Remington, The Science and Practice of Pharmacy, 22^(nd) Ed.,Philadelphia (2013).

As used herein the term “% by weight of a compound X based on the volumeof the composition”, also abbreviated as “% w/v”, corresponds to theamount of compound X in grams that is introduced in 100 mL of thecomposition.

As used herein the term “microparticle” refers to a particle having adiameter D₅₀ of 1 μm or greater to about 200 μm. The term “nanoparticle”refers to a particle having a diameter D₅₀ of less than 1 μm.

In exemplary embodiments, the diameter, which can be D₅₀, is 1 μm orgreater to about 200 μm; and the term “nanoparticle” refers to aparticle having a D₅₀ of less than about 1 μm.

As used herein an “ocular condition” is a disease, ailment or othercondition which affects or involves the eye, one of the parts or regionsof the eye, or the surrounding tissues such as the lacrimal glands.Broadly speaking, the eye includes the eyeball and the tissues andfluids which constitute the eyeball, the periocular muscles (such as theoblique and rectus muscles), the portion of the optic nerve which iswithin or adjacent to the eyeball and surrounding tissues such as thelacrimal glands and the eye lids.

As used herein an “anterior ocular condition” is a disease, ailment orcondition which affects or which involves an anterior (i.e. front of theeye) ocular region or site, such as a periocular muscle, an eye lid,lacrimal gland or an eye ball tissue or fluid which is located anteriorto the posterior wall of the lens capsule or ciliary muscles.

Thus, an anterior ocular condition primarily affects or involves one ormore of the following: the conjunctiva, the cornea, the anteriorchamber, the iris, the lens, or the lens capsule, and blood vessels andnerves which vascularize or innervate an anterior ocular region or site.An anterior ocular condition is also considered herein as extending tothe lacrimal apparatus. In particular, the lacrimal glands which secretetears, and their excretory ducts which convey tear fluid to the surfaceof the eye.

Moreover, an anterior ocular condition affects or involves the posteriorchamber, which is behind the retina but in front of the posterior wallof the lens capsule.

A “posterior ocular condition” is a disease, ailment or condition whichprimarily affects or involves a posterior ocular region or site such asthe choroid or sclera (in a position posterior to a plane through theposterior wall of the lens capsule), vitreous, vitreous chamber, retina,optic nerve (i.e. the optic disc), and blood vessels and nerves whichvascularize or innervate a posterior ocular region or site.

Thus, a posterior ocular condition can include a disease, ailment orcondition such as, for example, macular degeneration (such asnon-exudative age-related macular degeneration and exudative age-relatedmacular degeneration); choroidal neovascularization; acute macularneuroretinopathy; macular edema (such as cystoid macular edema anddiabetic macular edema); Behcet's disease, retinal disorders, diabeticretinopathy (including proliferative diabetic retinopathy); retinalarterial occlusive disease; central retinal vein occlusion; uveiticretinal disease; retinal detachment; ocular trauma which affects aposterior ocular site or location; a posterior ocular condition causedby or influenced by an ocular laser treatment; posterior ocularconditions caused by or influenced by a photodynamic therapy;photocoagulation; radiation retinopathy; epiretinal membrane disorders;branch retinal vein occlusion; anterior ischemic optic neuropathy;non-retinopathy diabetic retinal dysfunction, retinitis pigmentosa andglaucoma. Glaucoma can be considered a posterior ocular conditionbecause the therapeutic goal is to prevent the loss of or reduce theoccurrence of loss of vision due to damage to or loss of retinal cellsor optic nerve cells (i.e. neuroprotection).

An anterior ocular condition includes a disease, ailment or conditionsuch as, for example, aphakia; pseudophakia; astigmatism; blepharospasm;cataract; conjunctival diseases; conjunctivitis; corneal diseases;corneal ulcer; dry eye syndromes; eyelid diseases; lacrimal apparatusdiseases; lacrimal duct obstruction; myopia; presbyopia; pupildisorders; refractive disorders and strabismus. Glaucoma can also beconsidered to be an anterior ocular condition because a clinical goal ofglaucoma treatment can be to reduce a hypertension of aqueous fluid inthe anterior chamber of the eye (i.e. reduce intraocular pressure).

The present description is concerned with and directed to ophthalmiccompositions for topical drug delivery to the eye(s) and to methods forthe treatment of an ocular condition, such as an anterior ocularcondition or a posterior ocular condition or an ocular condition whichcan be characterized as both an anterior ocular condition and aposterior ocular condition.

Solid Complex of Cyclodextrin and Active Pharmaceutical Ingredient

The composition of the disclosure comprises a solid complex comprisingan active pharmaceutical ingredient and a cyclodextrin. The complexcomprising an active pharmaceutical ingredient and a cyclodextrin may bereferred to as an “active pharmaceutical ingredient/cyclodextrincomplex” or a “drug/cyclodextrin complex”. When the activepharmaceutical ingredient is dexamethasone and the cyclodextrin isγ-cyclodextrin, the complex comprising dexamethasone and γ-cyclodextrinmay be referred to as a “dexamethasone/γ-cyclodextrin complex”.

The solid complex of the composition of the disclosure may be a complexaggregate. The complex aggregate may correspond to an aggregate of aplurality of complexes, in particular a plurality of inclusion complexescomprising an active pharmaceutical ingredient and a cyclodextrin.

According to one embodiment, the ophthalmic composition of thedisclosure is a microsuspension. The term “microsuspension” is intendedto mean a composition comprising solid complex microparticles suspendedin a liquid phase.

In particular, the ophthalmic composition of the disclosure comprises asolid complex that has a diameter D₅₀ of less than about 100 μm, inparticular about 1 μm to about 100 μm. In one embodiment, the diameterD₅₀ may be in the range of about 1 μm to about 25 μm, in particularabout 1 μm to about 20 μm, more particularly about 1 μm to about 10 μm,even more particularly about 2 μm to about 10 μm, more particularlystill about 2 μm to about 5 μm or about 3 μm to about 8 μm. The diameterD₅₀ may be measured according to the test method described herein.

European Pharmacopoeia (01/2008:1163) teaches that eye drops in the formof a suspension should comply with the following: for each 10 μg ofsolid active substance, not more than about 20 particles have a maximumdimension greater than about 25 μm, and not more than about 2 of theseparticles have a maximum dimension greater than about 50 μm. None of theparticles can have a maximum dimension greater than about 90 μm. Thecompositions of the disclosure are in conformity with the requirementsof European Pharmacopoeia (01/2008:1163).

In general, it is recommended that particle sizes in aqueous eye dropsuspensions are kept to a minimum, preferable below about 10 μm, toprevent eye irritation. Furthermore, the sedimentation rate in aqueoussuspensions is proportional to the particle diameter, the sedimentationrate of large particles is faster than that of small particles assumingall other factors remaining constant.

Cyclodextrin

The composition of the disclosure comprises a cyclodextrin. Thecomposition of the disclosure may comprise a mixture of cyclodextrins.

Cyclodextrins, which are also known as cycloamyloses, are produced fromthe enzymatic conversion of starch. They have a cyclic structure that ishydrophobic on the inside and hydrophilic on the outside. Because of theamphiphilic nature of the ring, cyclodextrins have been known to enhancethe solubility and bioavailability of hydrophobic compounds.

As shown in FIG. 1 , cyclodextrins are cyclic oligosaccharidescontaining 6 (α-cyclodextrin), 7 (β-cyclodextrin), and 8(γ-cyclodextrin) glucopyranose monomers linked via α-1,4-glycosidebonds. α-Cyclodextrin, β-cyclodextrin and γ-cyclodextrin are naturalproducts formed by microbial degradation of starch. The outer surface ofthe doughnut shaped cyclodextrin molecules is hydrophilic, bearingnumerous hydroxyl groups, but their central cavity is somewhatlipophilic (Kurkov, S. V., Loftsson, T., 2013. Cyclodextrins. Int JPharm 453, 167-180; Loftsson, T., Brewster, M. E., 1996. Pharmaceuticalapplications of cyclodextrins. 1. Drug solubilization and stabilization.Journal of Pharmaceutical Sciences 85, 1017-1025). In addition to thethree natural cyclodextrins numerous water-soluble cyclodextrinderivatives have been synthesized and tested as drug carriers, includingcyclodextrin polymers (Stella, V. J., He, Q., 2008. Cyclodextrins. Tox.Pathol. 36, 30-42).

Cyclodextrins enhance the solubility and bioavailability of hydrophobiccompounds. In aqueous solutions, cyclodextrins form inclusion complexeswith many drugs by taking up a drug molecule, or more frequently somelipophilic moiety of the molecule, into the central cavity. Thisproperty has been used for drug formulation and drug delivery purposes.Formation of drug/cyclodextrin inclusion complexes, their effect on thephysicochemical properties of drugs, their effect on the ability ofdrugs to permeate biomembranes and the usage of cyclodextrins inpharmaceutical products have been reviewed (Loftsson, T., Brewster, M.E., 2010. Pharmaceutical applications of cyclodextrins: basic scienceand product development. Journal of Pharmacy and Pharmacology 62,1607-1621; Loftsson, T., Brewster, M. E., 2011. Pharmaceuticalapplications of cyclodextrins: effects on drug permeation throughbiological membranes.” J. Pharm. Pharmacol. 63, 1119-1135; Loftsson, T.,Järvinen, T., 1999. Cyclodextrins in ophthalmic drug delivery. AdvancedDrug Delivery Reviews 36, 59-79).

Cyclodextrins and drug/cyclodextrin complexes are able to self-assemblein aqueous solutions to form nano and micro-sized aggregates andmicellar-like structures that are also able to solubilize poorly solubleactive pharmaceutical ingredients through non-inclusion complexation andmicellar-like solubilization (Messner, M., Kurkov, S. V., Jansook, P.,Loftsson, T., 2010. Self-assembled cyclodextrin aggregates andnanoparticles. Int J Pharm 387, 199-208). In general, the tendency ofcyclodextrins to self-assemble and form aggregates increases uponformation of drug/cyclodextrin complexes and the aggregation increaseswith increasing concentration of active pharmaceuticalingredient/cyclodextrin complexes. In general, hydrophilic cyclodextrinderivatives, such as 2-hydroxypropyl-β-cyclodextrin and2-hydroxypropyl-γ-cyclodextrin, and their complexes are freely solublein water. On the other hand, the natural α-cyclodextrin, β-cyclodextrinand γ-cyclodextrin and their complexes have limited solubility in purewater or 129.5±0.7, 18.4±0.2 and 249.2±0.2 mg/ml, respectively, at 25°C. (Sabadini E., Cosgrovea T. and do Carmo Egidio F., 2006. Solubilityof cyclomaltooligosaccharides (cyclodextrins) in H₂O and D₂O: acomparative study. Carbohydr Res 341, 270-274). It is known that theirsolubility increases somewhat with increasing temperature (Jozwiakowski,M. J., Connors, K. A., 1985. Aqueous solubility behavior of threecyclodextrins. Carbohydr. Res., 143, 51-59). Due to the limitedsolubility of their complexes, the natural cyclodextrins most oftendisplay B_(s)-type or B_(i)-type phase-solubility diagrams (Brewster M.E., Loftsson T., 2007, Cyclodextrins as pharmaceutical solubilizers.Adv. Drug Deliv. Rev., 59, 645-666). It has been observed thatsolubility of the natural cyclodextrins can decrease below theirsolubility in pure water upon formation of active pharmaceuticalingredient/cyclodextrin complexes (FIG. 2 ) (Jansook, P., Moya-Ortega,M. D., Loftsson, T., 2010. Effect of self-aggregation of γ-cyclodextrinon drug solubilization. Journal of Inclusion Phenomena and MacrocyclicChemistry 68, 229-236). The low concentration of dissolved activepharmaceutical ingredient/cyclodextrin complexes hampers formation ofnano- and microparticles containing active pharmaceuticalingredient/cyclodextrin complexes. Furthermore, other excipients, suchas water-soluble polymers used to stabilize nano- and microsuspensions,can form complexes with cyclodextrins and, thus, hamper formation ofactive pharmaceutical ingredient/cyclodextrin complexes even further.

Previously, Applicants have described preparation and testing ofcyclodextrin-based eye drops containing dexamethasone (Johannesson, G.,Moya-Ortega, M. D., Asgrimsdottir, G. M., Lund, S. H., Thorsteinsdottir,M., Loftsson, T., Stefansson, E., 2014. Kinetics of γ-cyclodextrinnanoparticle suspension eye drops in tear fluid. Acta Ophthalmologica92, 550-556; Thorsteinn Loftsson and Einar Stefansson, Cyclodextrinnanotechnology for ophthalmic drug delivery, U.S. Pat. No. 7,893,040(Feb. 22, 2011); Thorsteinn Loftsson and Einar Stefansson, Cyclodextrinnanotechnology for ophthalmic drug delivery, U.S. Pat. No. 8,633,172(Jan. 21, 2014); Thorsteinn Loftsson and Einar Stefansson, Cyclodextrinnanotechnology for ophthalmic drug delivery U.S. Pat. No. 8,999,953(Apr. 7, 2015)), dorzolamide (Johannesson, G., Moya-Ortega, M. D.,Asgrimsdottir, G. M., Lund, S. H., Thorsteinsdottir, M., Loftsson, T.,Stefansson, E., 2014. Kinetics of γ-cyclodextrin nanoparticle suspensioneye drops in tear fluid. Acta Ophthalmologica 92, 550-556;Gudmundsdottir, B. S., Petursdottir, D., Asgrimsdottir, G. M.,Gottfredsdottir, M. S., Hardarson, S. H., Johannesson, G., Kurkov, S.V., Jansook, P., Loftsson, T., Stefansson, E., 2014. γ-Cyclodextrinnanoparticle eye drops with dorzolamide: effect on intraocular pressurein man. J. Ocul. Pharmacol. Ther. 30, 35-41), irbesartan (Muankaew, C.,Jansook, P., Stefansson, E., Loftsson, T., 2014. Effect ofγ-cyclodextrin on solubilization and complexation of irbesartan:influence of pH and excipients. Int J Pharm 474, 80-90), telmisartan (C.Muankaew, P. Jansook, H. H. Sigurõsson, T. Loftsson, 2016,Cyclodextrin-based telmisartan ophthalmic suspension: Formulationdevelopment for water-insoluble drugs. Int. J. Pharm. 507, 21-31) andcyclosporin A (S. Jóhannsdóttir, P. Jansook, E. Stefánsson, T. Loftsson,2015, Development of a cyclodextrin-based aqueous cyclosporin A eye dropformulation. Int. J. Pharm. 493(1-2), 86-95) in cyclodextrinnanoparticles. The studies show that the nanoparticles increase the drugcontact time with the ocular surface and the ocular bioavailability ofthe drugs. The active pharmaceutical ingredient/cyclodextrin nano- andmicroparticles are not only retained on the eye surface but also enhancedrug solubility in the aqueous tear fluid. Nano- and microparticlescomposed of active pharmaceutical ingredient/γ-cyclodextrin complexeshave been shown to be especially effective drug carriers for topicaldelivery of active pharmaceutical ingredient into the eye.

There are two approaches for preparation of nano- and microparticles andthe fabrication of nano and micro structures, the top-down approach andthe bottom-up approach. Top-down approach for preparation of activepharmaceutical ingredient/cyclodextrin nanoparticles and microparticlestypically involves milling of solid active pharmaceuticalingredient/cyclodextrin complexes to generate nanoparticles andmicroparticles of desired diameter. The top-down approach can introducesurface defects and contaminations. Bottom-up approach for preparationof active pharmaceutical ingredient/cyclodextrin nanoparticles impliesassembling of single molecules or single active pharmaceuticalingredient/cyclodextrin complexes into microparticles of desireddiameter. The bottom-up approach frequently leads to microparticlestructures with less defects and more homogeneous chemical composition.

Applicants have surprisingly discovered a bottom-up preparation ofdrug/cyclodextrin nanoparticles that can be accomplished with or withoutpresence of stabilizing polymers. According to the claimed method, adrug and cyclodextrin are suspended in an aqueous medium, such asaqueous eye drop medium, and heated. At a high temperature, the activecompounds as well as cyclodextrin and other pharmaceutical excipientsare fully or about fully dissolved in the aqueous media and theconcentration of drug/cyclodextrin complexes and excipient/cyclodextrincomplexes is much lower than at ambient temperature. Then the hotsolution is cooled at predetermined rate to promote formation ofparticles composed of drug/cyclodextrin complexes with diameter lessthan about 100 μm. The diameter of the particles can also be controlledby the heating and cooling cycle and by presence of stabilizing polymersin the aqueous medium. To prevent or substantially inhibit or reducedrug and/or excipient degradation, excessive heating of the medium isavoided followed by relatively rapid cooling to room temperature.

Under the controlled heating/cooling conditions, the aqueous solutioncomprising cyclodextrin is heated at a temperature and duration of timeto limit the formation of a degradation product, or sedimentation. Theinitial aqueous solution optionally further comprises an active agentdrug and/or a stabilizing polymer. During the heating cycle, the initialmilky white cyclodextrin solution is converted into a clear solution.Heating is accomplished by any method or means known to those havingordinary skill in the art. In preferred embodiments, the heating isaccomplished with an autoclave. For example, the autoclave can undergoabout a 20 to about 30 minute cycle at a temperature of from about 90°C. to about 120° C.

In an exemplary embodiment, under the controlled heating/coolingconditions, the aqueous solution comprising cyclodextrin is heated at atemperature and duration of time to limit the formation of a degradationproduct, or sedimentation. The initial aqueous solution optionallyfurther comprises an active pharmaceutical ingredient and/or astabilizing polymer. During the heating cycle, the initial milky whitecyclodextrin solution is converted into a clear solution. Heating isaccomplished by any method or means known to those having ordinary skillin the art. In preferred embodiments, the heating is accomplished withan autoclave or jacketed reactors with steam. For example, the autoclavecan undergo about a 10 to about 30 minute cycle at a temperature ofabout 121° C.

The heated solution is then cooled at a sufficient rate to produce adrug/cyclodextrin complex having a diameter of less than about 100 μm.Upon cooling, the drug/cyclodextrin complex precipitates to form thedesired microsuspension. The microsuspension comprises about 70% toabout 99% of the drug in microparticles and about 1% to about 30% of thedrug in nanoparticles the eye drop vehicle. The microparticles have anaverage diameter of about 1 μm to about 100 μm. It is possible for theaverage diameter of microparticles to be in the range of about 1 μm toabout 20 μm, about 1 μm to about 25 μm, about 1 μm to about 10 μm, orabout 2 μm to about 5 μm. In an exemplary embodiment, themicrosuspension comprises about 80% of the drug to be in microparticleshaving an average diameter of about 1 μm to about 10 μm, and about 20%of the drug to be in nanoparticles.

In one embodiment, the heated solution is then cooled at a sufficientrate to produce drug/cyclodextrin complex aggregates having a diameterof less than about 100 μm. Upon cooling, the drug/cyclodextrin complexprecipitates to form the desired microsuspension. The microsuspensioncomprises about 40% to about 99% of the drug in microparticles and about1% to about 60% of the drug in nanoparticles or water-solubledrug/cyclodextrin complexes. The microparticles have an average diameterof about 1 μm to about 100 μm. It is possible for the average diameterof microparticles to be in the range of about 1 μm to about 20 μm, about1 μm to about 25 μm, about 1 μm to about 10 μm, or about 2 μm to about 5μm. In an exemplary embodiment, the microsuspension comprises about 80%of the drug to be in microparticles having an average diameter of about1 μm to about 10 μm, and about 20% of the drug to be in nanoparticles.

The microsuspensions prepared according to the claimed procedure haveabout a 10-fold to 100-fold increase in dissolved active agent drugconcentration when compared to microsuspensions prepared according toknown methods in the form of water-soluble nanoparticles, individualdrug/cyclodextrin complexes and dissolved drug molecules. For example,known dexamethasone compositions comprise a dexamethasone concentrationof about 1 mg/mL where only 0.1 mg/mL is in solution. However, adexamethasone/cyclodextrin composition prepared according to the claimedmethod can comprise a dexamethasone concentration of about 15 mg/mLwhere about 4 mg/mL is in solution.

In a preferred embodiment, the cyclodextrin is α-cyclodextrin,β-cyclodextrin, γ-cyclodextrin, or combinations thereof.

In a particularly preferred embodiment, the cyclodextrin isγ-cyclodextrin. Indeed, γ-cyclodextrin has a higher solubility in watercompared to that of α-cyclodextrin and β-cyclodextrin. Moreover,γ-cyclodextrin is prone to hydrolysis into glucose and maltose subunitsby α-amylase in the tear fluid and the gastrointestinal tract.

The amount of cyclodextrin in the ophthalmic composition of thedisclosure may be from 1 to 25%, in particular 5 to 20%, moreparticularly 10 to 18%, even more particularly 12 to 16%, by weight, ofcyclodextrin based on the volume of the composition.

In addition to the cyclodextrin, the ophthalmic composition of thedisclosure may further comprise a water-soluble cyclodextrin derivativeselected from the group consisting of 2-hydroxypropyl-α-cyclodextrin,2-hydroxypropyl-1-cyclodextrin, 2-hydroxypropyl-γ-cyclodextrin,sulfobutyl ether α-cyclodextrin, sulfobutyl ether β-cyclodextrin,sulfobutyl ether γ-cyclodextrin, methylated α-cyclodextrin, methylatedβ-cyclodextrin, methylated γ-cyclodextrin, and combinations thereof. Thewater-soluble cyclodextrin derivative may especially be used to furtherenhance the solubility of the active pharmaceutical ingredient, i.e. theamount of active pharmaceutical ingredient that is dissolved in thecomposition.

Active Pharmaceutical Ingredient

The composition of the disclosure comprises an active pharmaceuticalingredient.

The active pharmaceutical ingredient may be referred to as a “drug”. Inthe context of the disclosure, the active pharmaceutical ingredient isan ophthalmic drug, i.e. a compound that exhibits a therapeutic effectwhen administered in a sufficient amount to a patient suffering from anocular condition.

In particular, the ophthalmic composition may comprise an activepharmaceutical ingredient selected from the group consisting of asteroid such as dexamethasone, difluprednate, estradiol, fluocinolone,fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone,triamcinolone, and rimexolone; a kinase inhibitor such as axitinib,BMS-794833(N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide),carbozantinib, cediranib, crizotinib, dasatinib, dovitinib, everolimus,lapatinib, lenvatinib, motesanib, nilotinib, nintedanib, orantinib,PD173074(N-[2-[[4-(Diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]-N′-(1,1-dimethylethyl)urea),pazopanib, regorafenib, ruxolitinib, sorafenib, sunitinib, temsirolimus,tofacitinib, vandetanib, vemurafenib, and ZM323881(5-((7-Benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol); anangiotensin II receptor antagonist such as candesartan, irbesartan,losartan, olmesartan, telmisartan, and valsartan; an aldose reductaseinhibitor such as 2-methylsorbinol; an immunosuppressant such assirolimus; a carbonic anhydrase inhibitor such as acetazolamide,brinzolamide, dorzolamide, ethoxzolamide and methazolamide; anantimicrobial or an antiviral such as acyclovir, chloramphenicol,chlortetracycline, ciprofloxacin, fusidic acid, gancyclovir,norfloxacin, ofloxacin, tetracycline, and zidovudine; an antihistaminesuch as levocabastine; and a non-steroidal anti-inflammatory activepharmaceutical ingredient such as bromfenac, diclofenac, indomethacinand nepafenac; and combinations thereof.

The active agent drug for use in the nano- and microparticles in theexemplary embodiments can be selected from, but are not limited to, thegroup consisting of a steroid such as dexamethasone, difluprednate,estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednoletabonate, prednisolone, triamcinolone and rimexolone; a kinaseinhibitor such as axitinib, BMS-794833N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide),carbozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib,nintedanib, orantinib, PD173074(N-[2-[[4-(Diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]-N′-(1,1-dimethylethyl)urea),pazopanib, regorafenib, sorafenib, tofacitinib, and ZM323881(5-((7-Benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol); anangiotensin II receptor antagonist such as candesartan, irbesartan,losartan, olmesartan, telmisartan, and valsartan; an aldose reductaseinhibitors such as 2-methylsorbinol; an immunosuppressant such assirolimus, a carbonic anhydrase inhibitor such as acetazolamide,brinzolamide, dorzolamide, ethoxzolamide and methazolamide; anantimicrobial and an antiviral such as acyclovir, chloramphenicol,chlortetracycline, ciprofloxacin, fusidic acid, gancyclovir,norfloxacin, ofloxacin, tetracycline, and zidovudine; an antihistaminesuch as levocabastine; and a non-steroidal anti-inflammatory drug suchas bromfenac, diclofenac, indomethacin, nepafenac, or a combinationthereof.

According to a preferred embodiment, the ophthalmic compositioncomprises an active pharmaceutical ingredient selected from the groupconsisting of dexamethasone, axitinib, cediranib, dovitinib, motesanib,pazopanib, regorafenib, losartan, olmesartan, dorzolamide, diclofenac,nepafenac, and combinations thereof. More preferably, the activepharmaceutical ingredient is dexamethasone.

The concentration of active pharmaceutical ingredient in the ophthalmiccomposition of the disclosure may be from about 0.1 mg/mL to about 100mg/mL, in particular from about 1 mg/mL to about 100 mg/mL, inparticular from about 1 mg/mL to about 50 mg/mL, more particularly fromabout 1 mg/mL to about 20 mg/mL, even more particularly about 5 mg/mL toabout 25 mg/mL, more particularly still from about 10 mg/mL to about 20mg/mL.

In exemplary embodiments, the active pharmaceutical ingredient (i.e.drug) is present in the initial aqueous solution at a concentration ofabout 1 mg/mL to about 100 mg/mL. It is further possible to obtain thedesired final active pharmaceutical ingredient/cyclodextrin complexconcentration with an initial active pharmaceutical ingredientconcentration of about 1 mg/mL to about 50 mg/mL and about 1 mg/mL toabout 20 mg/mL.

In other exemplary embodiments, the active pharmaceutical ingredient ispresent in the initial aqueous solution at a concentration of about 0.01mg/mL to about 10 mg/m L.

The compositions of the disclosure may have about 10-fold to about100-fold increase in dissolved active pharmaceutical ingredientconcentration when compared to compositions prepared according to knownmethods.

In particular, 60 to 95% by weight, more particularly 70 to 90% byweight, of the active pharmaceutical ingredient in the composition maybe in the form of a solid complex of active pharmaceutical ingredientand cyclodextrin.

Even more particularly, 5 to 40% by weight, in particular 10 to 30% byweight, of the active pharmaceutical ingredient in the composition maybe in dissolved form. The dissolved form includes uncomplexed activepharmaceutical ingredient that is dissolved in the liquid phase andcomplexes of active pharmaceutical ingredient and cyclodextrin that aredissolved in the liquid phase as well as water-soluble nanoparticlesconsisting of drug/cyclodextrin complex aggregates.

Preferably, 0% to 0.5% by weight of the active pharmaceutical ingredientin the composition may be in uncomplexed solid form. As such, thecomposition of the disclosure may be substantially free of soliduncomplexed particles of active pharmaceutical ingredient.

In one embodiment, the microsuspension may comprise about 70% to about99% of the active pharmaceutical ingredient in microparticles and about1% to about 30% of the active pharmaceutical ingredient innanoparticles. More particularly, the microsuspension may comprise about80% of the active pharmaceutical ingredient in microparticles having adiameter of about 1 μm to about 10 μm, and about 20% of the activepharmaceutical ingredient in nanoparticles.

In another embodiment, the microsuspension may comprise about 40% toabout 99% of the active pharmaceutical ingredient in microparticles andabout 1% to about 60% of the active pharmaceutical ingredient innanoparticles or water-soluble active pharmaceuticalingredient/cyclodextrin complexes. In particular, the microsuspensionmay comprise about 80% to about 90% of the active pharmaceuticalingredient in microparticles having a diameter of about 1 μm to about 10μm, and about 10% to about 20% of the active pharmaceutical ingredientin nanoparticles or water-soluble active pharmaceuticalingredient/cyclodextrin complexes.

Polymer

The composition of the disclosure may further comprise a polymer.

In particular, said polymer may be a water-soluble polymer. Moreover,said polymer may be a viscosity enhancing polymer. The term “viscosityenhancing polymer” is intended to mean a polymer that increases theviscosity of a liquid. The polymer increases the viscosity of thecomposition of the disclosure. The increase of viscosity results is aenhanced physical stability of the composition. As such, the compositionis less prone to sedimentation of the solid complex when it comprises apolymer. The polymer may thus be considered as a polymeric stabilizingagent.

In particular, the polymer may be a surface active polymer. The term“surface active polymer” is intended to mean a polymer that exhibitssurfactant properties. Surface active polymers may, for example,comprise hydrophobic chains grafted to a hydrophilic backbone polymer;hydrophilic chains grafted to a hydrophobic backbone; or alternatinghydrophilic and hydrophobic segments. The first two types are calledgraft copolymers and the third type is named block copolymer.

In one embodiment, the ophthalmic composition of the disclosurecomprises a polymer selected from the group consisting of apolyoxyethylene fatty acid ester; a polyoxyethylene alkylphenyl ether; apolyoxyethylene alkyl ether; a cellulose derivative such as alkylcellulose, hydroxyalkyl cellulose and hydroxyalkyl alkylcellulose; acarboxyvinyl polymer such as a carbomer, for example Carbopol 971 andCarbopol 974; a polyvinyl polymer; a polyvinyl alcohol; apolyvinylpyrrolidone; a copolymer of polyoxypropylene andpolyoxyethylene; tyloxapol; and combinations thereof.

Examples of suitable polymers include, but are not limited to,polyethylene glycol monostearate, polyethylene glycol distearate,hydroxypropyl methylcellulose, hydroxypropylcellulose,polyvinylpyrrolidone, polyoxyethylene lauryl ether, polyoxyethyleneoctyldodecyl ether, polyoxyethylene stearyl ether, polyoxyethylenemyristyl ether, polyoxyethylene oleyl ether, sorbitan esters,polyoxyethylene hexadecyl ether (e.g., cetomacrogol 1000),polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fattyacid esters (e.g., Tween 20 and Tween 80 (ICI Specialty Chemicals));polyethylene glycols (e.g., Carbowax 3550 and 934 (Union Carbide)),polyoxyethylene stearates, carboxymethylcellulose calcium,carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose,hydroxypropyl methylcellulose, cellulose, polyvinyl alcohol (PVA),poloxamers (e.g., Pluronics F68 and FI08, which are block copolymers ofethylene oxide and propylene oxide); poloxamines (e.g., Tetronic 908,also known as Poloxamine 908, which is a tetrafunctional block copolymerderived from sequential addition of propylene oxide and ethylene oxideto ethylenediamine (BASF Wyandotte Corporation, Parsippany, N.J.));Tetronic 1508 (T-1508) (BASF Wyandotte Corporation), Tritons X-200,which is an alkyl aryl polyether sulfonate (Rohm and Haas);PEG-derivatized phospholipid, PEG-derivatized cholesterol,PEG-derivatized cholesterol derivative, PEG-derivatized vitamin A,PEG-derivatized vitamin E, random copolymers of vinyl pyrrolidone andvinyl acetate, combinations thereof and the like.

Particularly preferred examples of polymers according to the disclosureare tyloxapol and a copolymer of polyoxypropylene and polyoxyethylene.

More particularly, the copolymer of polyoxypropylene and polyoxyethylenemay be a triblock copolymer comprising a hydrophilic block-hydrophobicblock-hydrophilic block configuration.

In one embodiment, the composition of the disclosure comprises a polymerwhich is a poloxamer. Poloxamers can include any type of poloxamer knownin the art. Poloxamers include poloxamer 101, poloxamer 105, poloxamer108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181,poloxamer 182, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231,poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333,poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer402, poloxamer 403, poloxamer 407, poloxamer 105 benzoate and poloxamer182 dibenzoate. Poloxamers are also referred to by their trade namePluronic such as Pluronic 10R5, Pluronic 17R2, Pluronic 17R4, Pluronic25R2, Pluronic 25R4, Pluronic 31R1, Pluronic F 108, Pluronic F 108,Pluronic F 108, Pluronic F 108NF, Pluronic F 127, Pluronic F 127 NF,Pluronic F 127, Pluronic F 127, Pluronic F 38, Pluronic F 38, Pluronic F68, Pluronic F 77, Pluronic F 87, Pluronic F 88, Pluronic F 98, PluronicL 10, Pluronic L 101, Pluronic L 121, Pluronic L 31, Pluronic L 35,Pluronic L 43, Pluronic L 44, Pluronic L 61, Pluronic L 62, Pluronic L62 LF, Pluronic L 62D, Pluronic L 64, Pluronic L 81, Pluronic L 92,Pluronic L 44, Pluronic N 3, Pluronic P 103, Pluronic P 104, Pluronic P105, Pluronic P 123, Pluronic P 65, Pluronic P 84, Pluronic P 85,combinations thereof and the like.

Especially useful polymers as stabilizers are poloxamers. Poloxamers caninclude any type of poloxamer known in the art. Poloxamers includepoloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 183,poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235,poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335,poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, poloxamer407, poloxamer 105 benzoate and poloxamer 182 dibenzoate. Poloxamers arealso referred to by their trade name Pluronic such as Pluronic 10R5,Pluronic 17R2, Pluronic 17R4, Pluronic 25R2, Pluronic 25R4, Pluronic31R1, Pluronic F 108 Cast Solid Surfacta, Pluronic F 108 NF, Pluronic F108 Pastille, Pluronic F 108NF Prill Poloxamer 338, Pluronic F 127,Pluronic F 127 NF, Pluronic F 127 NF 500 BHT Prill, Pluronic F 127 NFPrill Poloxamer 407, Pluronic F 38, Pluronic F 38 Pastille, Pluronic F68, Pluronic F 68 Pastille, Pluronic F 68 LF Pastille, Pluronic F 68 NF,Pluronic F 68 NF Prill Poloxamer 188, Pluronic F 77, Pluronic F 77Micropastille, Pluronic F 87, Pluronic F 87 NF, Pluronic F 87 NF PrillPoloxamer 237, Pluronic F 88, Pluronic F 88 Pastille, Pluronic F 98,Pluronic L 10, Pluronic L 101, Pluronic L 121, Pluronic L 31, Pluronic L35, Pluronic L 43, Pluronic L 44 NF Poloxamer 124, Pluronic L 61,Pluronic L 62, Pluronic L 62 LF, Pluronic L 62D, Pluronic L 64, PluronicL 81, Pluronic L 92, Pluronic L44 NF INH surfactant Poloxamer 124 View,Pluronic N 3, Pluronic P 103, Pluronic P 104, Pluronic P 105, Pluronic P123 Surfactant, Pluronic P 65, Pluronic P 84, Pluronic P 85,combinations thereof and the like.

A further polymeric stabilizing agent compatible with the compositionsand methods described herein is tyloxapol. In preferred embodiments, thestabilizer and co-solubilizer is tyloxapol, which is a4-(1,1,3,3-tetramethylbutyl)phenol polymer with formaldehyde andoxirane.

The solutions and microsuspensions prepared according to Applicants'method optionally comprise further additives. For example, it isenvisioned that the solution and/or the microsuspension furthercomprises ethylenediaminetetraacetic acid (EDTA). EDTA can be used, forexample, to reduce degradation or as a stabilizer. It is also envisionedthat the solution and/or microsuspension is isotonic, for example, withthe addition of sodium chloride.

In an exemplary embodiment, the EDTA can be ethylenediaminetetraaceticacid disodium salt.

According to one method, the active agent drug and at least onecyclodextrin are suspended in an aqueous eye drop vehicle to provide asuspension having a milky appearance. The suspension is then heated fora sufficient time at a sufficient temperature until both the drug andcyclodextrin are dissolved in the aqueous eye drop solution, and nodegradation product is formed. Once the drug and the cyclodextrin aredissolved the milky suspension turns into a substantially clearsolution. The resulting solution is cooled at a rate sufficient toproduce a microsuspension comprising a microparticle drug/cyclodextrincomplex.

In another embodiment, an active agent drug, at least one cyclodextrin,and at least one polymer are suspended in an aqueous eye drop vehicle toprovide a suspension having a milky appearance. The suspension is thenheated for a sufficient time at a sufficient temperature until the drug,cyclodextrin, and the polymer are dissolved in the aqueous eye dropsolution, and no degradation product is formed. Once the drug, thecyclodextrin, and the polymer are dissolved the milky suspension turnsinto a substantially clear solution. The resulting solution is cooled ata rate sufficient to produce a microsuspension comprising amicroparticle drug/cyclodextrin/polymer complex. In this alternativeembodiment, drug/cyclodextrin/polymer complex comprises a polymercoating.

In an alternative method, at least one cyclodextrin is suspended in anaqueous eye drop vehicle to provide a suspension having a milkyappearance. The cyclodextrin suspension is heated for a sufficient timeat a sufficient temperature until the cyclodextrin is dissolved in theaqueous eye drop solution. An active agent drug is added to the heatedaqueous suspension, while stirring the solution, until the drug isdissolved in the solution. The resulting solution is cooled at a ratesufficient to produce a microsuspension comprising a microparticledrug/cyclodextrin complex.

In yet another method, cyclodextrin and at least one polymer aresuspended in an aqueous eye drop vehicle to provide a suspension havinga milky appearance. Once the cyclodextrin and the polymer are dissolvedthe milky suspension turns into a substantially clear solution. Anactive agent drug is added to the heated aqueous suspension, whilestirring the solution, until the drug is dissolved in the solution. Theresulting solution is cooled at a rate sufficient to produce amicrosuspension comprising a microparticle drug/cyclodextrin/polymercomplex. The resulting drug/cyclodextrin/polymer complex comprises apolymer coating.

In another method, at least one polymer and a drug is suspended in anaqueous eye drop vehicle to provide a suspension having a milkyappearance. In another container, at least one cyclodextrin is suspendedin water to provide a suspension having a milky appearance. Both thesuspensions are heated for a sufficient time at a sufficient temperatureuntil the cyclodextrin solution becomes transparent, the polymer/drugsuspension is still a milky suspension and no (or substantially no)degradation product is being formed. The cyclodextrin solution is addedto the polymer/drug phase and the mixture becomes clear as the drugdissolves and the solution is mixed for a sufficient time at the sametemperature. The resulting solution is cooled at a rate sufficient toproduce a microsuspension comprising a microparticle drug/cyclodextrincomplex.

The polymer that may be introduced in the composition of the disclosurecan exhibit a weight average molecular weight of 2,000 g/mol or higher,in particular a weight average molecular weight from 2,000 to 50,000g/mol, more particularly 5,000 to 25,000 g/mol, even more particularly9,000 to 15,000 g/mol.

The amount of polymer in the composition of the disclosure may be 0.5 to5%, in particular 1 to 4%, more particularly 2 to 3%, more particularly2.2 to 2.8%, by weight of polymer based on the volume of thecomposition.

When the composition comprises a polymer, the viscosity of thecomposition may be from 4 to 14 cP, preferably 5 to 13 cP, morepreferably 6 to 12 cP.

Part of the polymer that is introduced in the composition of thedisclosure may be contained in the solid complexes of activepharmaceutical ingredient and cyclodextrin. As such, some of the polymermay be taken up within the solid complex and/or part of the polymer maybe coated on the surface of the solid complex. The microsuspension maythus comprise a microparticle drug/cyclodextrin/polymer complex. Saiddrug/cyclodextrin/polymer complex may comprise a polymer coating.

Ophthalmically Acceptable Medium

The composition of the disclosure comprises an ophthalmically acceptablemedium.

The term “ophthalmically acceptable medium” is intended to mean a mediumsuitable for ophthalmic administration of the composition. Theophthalmically acceptable medium is preferably a liquid. Theophthalmically acceptable medium may notably comprise water. Inparticular, the ophthalmically acceptable medium does not comprise anyother solvent than water. The ophthalmically acceptable medium may thuscorrespond to an aqueous eye drop vehicle.

According to a preferred embodiment the ophthalmically acceptable mediumcomprises water and optionally an additive selected from the groupconsisting of a preservative, a stabilizing agent, an electrolyte, abuffering agent, and combinations thereof.

In particular, the ophthalmically acceptable medium may comprise apreservative. A preservative may be used to limit bacterialproliferation in the composition.

Suitable examples of preservative are sodium bisulfite, sodiumbisulfate, sodium thiosulfate, benzalkonium chloride, chlorobutanol,thimerosal, phenylmercuric acetate, phenylmercuric nitrate,methylparaben, phenylethyl alcohol, and combinations thereof.Preferably, the preservative is benzalkonium chloride.

The amount of preservative in the composition of the disclosure may be 0to 1%, in particular 0.001 to 0.5%, more particularly 0.005 to 0.1%,even more particularly 0.01 to 0.04%, by weight of preservative based onthe volume of the composition.

In particular, the ophthalmically acceptable medium may comprise astabilizing agent. A stabilizing agent may be used to reduce degradationor stabilize the composition during storage.

An example of a suitable stabilizing agent is disodium edetate.

The amount of stabilizing agent in the composition of the disclosure maybe 0 to 1%, in particular 0.01 to 0.5%, more particularly 0.08 to 0.2%by weight of stabilizing agent based on the volume of the composition.

In particular, the ophthalmically acceptable medium may comprise anelectrolyte. An electrolyte may especially be used to make thecomposition isotonic.

Examples of suitable electrolytes include sodium chloride, potassiumchloride, and combinations thereof. Preferably, the electrolyte issodium chloride.

The amount of electrolyte in the composition of the disclosure may be 0to 2%, in particular 0.1 to 1.5%, more particularly 0.5 to 1% by weightof electrolyte based on the volume of the composition.

Impurities

The composition according to the disclosure may notably exhibit a lowconcentration of impurities. The low amount of impurities in thecomposition of the disclosure results from the specific preparationprocess described hereinafter, in particular the specific heating andcooling steps implemented in the preparation process.

Accordingly, the composition according to the disclosure may compriseless than 2%, in particular less than 1%, more particularly less than0.8%, by weight of impurities based on the weight of the activepharmaceutical ingredient.

The term “impurities” is intended to mean a product that was notvoluntarily introduced in the composition of the disclosure but wasgenerated in situ during manufacturing of the composition. As such theterm “impurities” encompasses any product other than an activepharmaceutical ingredient, a cyclodextrin, a complex of activepharmaceutical ingredient and a cyclodextrin, a polymer, water, apreservative, a stabilizing agent, and an electrolyte as defined hereinabove. The impurities typically correspond to a by-product or adegradation product of the active pharmaceutical ingredient. The amountof the impurities in the composition can be determined by conventionalanalytical techniques including, for example liquid chromatography, massspectrometry and/or NMR. The amount of impurities can be measuredshortly after, for example less than 24 hours after, preparation of thecomposition or after storage of the composition, for example up to 2years of storage of the composition, at 25° C.

Surprisingly, Applicants observed that the compositions of thedisclosure containing specific active pharmaceutical ingredients wereparticularly prone to generating impurities when aqueous solutions orsuspensions thereof were heated in the presence of cyclodextrins, forexample γ-cyclodextrin, at a temperature above 120° C. However,negligible amounts of impurities were obtained when the compositionswere prepared according to the method of the disclosure describedhereinafter.

Accordingly, according to one embodiment, the present disclosureprovides an ophthalmic composition comprising, in an ophthalmicallyacceptable medium, a solid complex comprising an active pharmaceuticalingredient and a cyclodextrin; wherein the composition comprises lessthan 2%, in particular less than 1%, more particularly less than 0.8%,by weight of impurities based on the weight of the active pharmaceuticalingredient; wherein the active pharmaceutical ingredient is selectedfrom the group consisting of dexamethasone, axitinib, cediranib,dovitinib, motesanib, pazopanib, regorafenib, losartan, olmesartan,dorzolamide, diclofenac, nepafenac, and combinations thereof; andwherein the cyclodextrin is γ-cyclodextrin.

In particular, when the active ingredient is dexamethasone, thecomposition of the disclosure may comprise less than 0.5%, in particularless than 0.3%, more particularly less than 0.2%, by dexamethasone enolaldehydes (i.e. a mixture of Z and E isomers) based on the weight ofdexamethasone.

Dexamethasone enol aldehydes are dehydrated dexamethasone that have thefollowing structures:

Furthermore, when the active ingredient is dexamethasone, thecomposition of the disclosure may comprise less than 0.5%, in particularless than 0.3%, more particularly less than 0.2%, by weight of16,17-unsaturated dexamethasone based on the weight of dexamethasone.

16,17-unsaturated dexamethasone is a dehydrated dexamethasone that hasthe following structure:

In particular, when the active ingredient is dexamethasone, thecomposition of the disclosure may comprise less than 0.5%, in particularless than 0.3%, more particularly less than 0.2%, by weight ofdehydrated dexamethasone, i.e. dexamethasone enol aldehydes (i.e. amixture of Z and E isomers) and 16,17-unsaturated dexamethasone, basedon the weight of the dexamethasone.

It is known that dexamethasone can undergo base catalyzed andphotochemical degradation in aqueous solutions (E. M. Cohen, 1973,Dexamethasone. Analytical Profiles of Drug Substances, 2, 163-197) andthat the active pharmaceutical ingredient is subjected to oxidativedecomposition (R. E. Conrow, G. W. Dillow, L. Bian, L. Xue, O.Papadopoulou, J. K. Baker, B. S. Scott, 2002, Corticosteroiddecomposition via a mixed anhydride. J. Org. Chem. 67, 6835-6836). Thedexamethasone monograph in the European Pharmacopoeia (01/2014:0388)lists 11 impurities and degradation products and describes a method fortheir detection. The British Pharmacopeia (2015, version 19.0) has amonograph on dexamethasone eye drop suspension and lists 5 degradationimpurities and products and a method for their detection. For maximumchemical stability in aqueous solutions the pH of the dexamethasone eyedrop suspensions should be kept between about 5.0 and about 6.0. Themajor degradation product formed during preparation of the aqueousdexamethasone eye drop suspension containing cyclodextrin andsterilization of the eye drops in an autoclave is believed to comprise a16,17-unsaturated dexamethasone and a mixture of E and Z isomers ofdexamethasone enol aldehydes, formed by Mattox rearrangement through thedehydration of dexamethasone (B. Chen, M. Li, M. Lin, G. Tumambac, A.Rustum, 2009, A comparative study of enol aldehyde formation frombetamethasone, dexamethasone, beclomethasone and related compound underacidic and alkaline conditions. Steroids, 74, 30-41). Due to sterichindrance in the case of dexamethasone the main product is believed tobe the dexamethasone enol aldehyde Z isomer. This degradation product isnot listed in the pharmacopoeias. Previously, 16,17-unsaturateddexamethasone has been detected in parenteral dexamethasone solutionsthat were heated to about 75° C. for about 10 days (M. Spangler, E.Mularz, 2001, A validated, stability-indicating method for the assay ofdexamethasone in drug substance and drug product analyses, and the assayof preservatives in drug product. Chromatographia, 54, 329-334), whichis also a dehydrated degradation product of dexamethasone and since theauthors did not analyze the product the authors likely detected the enolaldehydes. Previously a Japanese group described the two enol aldehydesand the 16-17 unsaturated degradation products of betamethasone, both byacidic catalysis (T. Hidaka, S. Huruumi, S. Tamaki, M. Shiraishi, H.Minato, 1980, Studies on betamethasone: behavior of betamethasone inacid or alkaline medium, photolysis and oxidation. Yakugaku Zasshi, 100,72-80). The apparent activation energy for the rate of dexamethasonedegradation to form 16,17-unsaturated dexamethasone and dexamethasoneenol aldehydes in aqueous γ-cyclodextrin solution is unusually high and,thus, these degradation products are essentially not formed at ambienttemperature. The presence of cyclodextrin in the aqueous eye dropsappears to promote dehydration of dexamethasone during autoclaving toform 16,17-unsaturated dexamethasone and a mixture of E and Z isomers ofdexamethasone enol aldehydes.

In aqueous solutions, diclofenac is relatively stable at roomtemperature when protected from light and oxygen (R. Chadaha, N. Kashid,D. V. S. Jain, 2003, Kinetics of degradation of diclofenac sodium inaqueous solution determined by a calorimetric method. Pharmazie, 58,631-635). Although β-cyclodextrin has been shown to stabilize diclofenacin aqueous solutions at about pH 7 we have observed that γ-cyclodextrincan accelerate the degradation during autoclaving causing intensecoloring of aqueous diclofenac solutions.

Here, Applicants have surprisingly discovered that the claimed methodprovides solutions and microsuspensions comprising active pharmaceuticalingredient/cyclodextrin complexes that are stable in the aqueoussolution. For example, the claimed method provides adexamethasone/γ-cyclodextrin eye drop solution in which very low amountsof 16,17-unsaturated dexamethasone and dexamethasone enol aldehydes areformed. Moreover, this disclosure provides a diclofenac/γ-cyclodextrinsolution in which no degradation product or sedimentation is observedfor at least about 12 months. These aqueous γ-cyclodextrin containingeye drops also have the benefit of having 10 to 100-fold increase, inthe case of dexamethasone about a 30-fold increase, in the concentrationof dissolved active pharmaceutical ingredient, and have the desiredparticle size to achieve maximum drug diffusion.

Dexamethasone Composition

According to a particularly preferred embodiment, the present disclosureprovides an ophthalmic dexamethasone composition comprising, in anophthalmically acceptable medium, a solid complex comprisingdexamethasone and γ-cyclodextrin.

In one embodiment, the ophthalmic dexamethasone composition of thedisclosure comprises less than 0.5%, in particular less than 0.3%, moreparticularly less than 0.2%, by weight of 16,17-unsaturateddexamethasone and dexamethasone enol aldehydes based on the weight ofdexamethasone.

In another embodiment, the ophthalmic dexamethasone composition of thedisclosure comprises a polymer as defined herein above. The viscosity ofsaid ophthalmic dexamethasone composition may be from 4 to 14 cP,preferably 5 to 13 cP, more preferably 6 to 12 cP.

In yet another embodiment, the ophthalmic dexamethasone composition ofthe disclosure comprises less than 0.5%, in particular less than 0.3%,more particularly less than 0.2%, by weight of 16,17-unsaturateddexamethasone and dexamethasone enol aldehydes based on the weight ofdexamethasone; the ophthalmic dexamethasone composition of thedisclosure comprises a polymer; and the viscosity of the ophthalmicdexamethasone composition is from 4 to 14 cP, preferably 5 to 13 cP,more preferably 6 to 12 cP.

The concentration of dexamethasone in the ophthalmic composition of thedisclosure may be from 10 mg/mL to 20 mg/mL. As such, the amount ofdexamethasone in the composition of the disclosure is much higher thanknown dexamethasone compositions which comprise a dexamethasoneconcentration of about 1 mg/mL where about 0.1 mg/mL is in solution. Inparticular, the concentration of dexamethasone in the ophthalmiccomposition of the disclosure may be of about 15 mg/mL where about 4mg/mL is in solution.

In particular, 60 to 95% by weight, more particularly 70 to 90% byweight, of the dexamethasone in the composition may be in the form of asolid complex of dexamethasone and γ-cyclodextrin.

More particularly, 5 to 40% by weight, in particular 10 to 30% byweight, of the dexamethasone in the composition may be in dissolvedform. The dissolved form includes uncomplexed dexamethasone that isdissolved in the liquid phase, complexes of dexamethasone andcyclodextrin that are dissolved in the liquid phase and water-solublenanoparticles consisting of dexamethasone/cyclodextrin complexaggregates.

Preferably, 0% to 0.5% by weight of the dexamethasone in the compositionmay be in uncomplexed solid form. As such, the composition of thedisclosure may be substantially free of solid uncomplexed particles ofdexamethasone.

In one embodiment, the microsuspension may comprise about 70% to about99% of the dexamethasone in microparticles and about 1% to about 30% ofthe dexamethasone in nanoparticles. More particularly, themicrosuspension may comprise about 80% of the dexamethasone inmicroparticles having a diameter of about 1 μm to about 10 μm, and about20% of the dexamethasone in nanoparticles.

In another embodiment, the microsuspension may comprise 40% to 99% ofthe dexamethasone in microparticles and about 1% to about 60% of thedexamethasone in nanoparticles or water-solubledexamethasone/γ-cyclodextrin complexes. In particular, themicrosuspension may comprise about 80 to 90% of the dexamethasone inmicroparticles having a diameter of about 1 μm to about 10 μm, and about10 to 20% of the dexamethasone in nanoparticles or water-solubledexamethasone/γ-cyclodextrin complexes.

The amount of γ-cyclodextrin in the ophthalmic dexamethasone compositionmay be from 1 to 25%, in particular 5 to 20%, more particularly 10 to18%, even more particularly 12 to 16%, by weight of γ-cyclodextrin basedon the volume of the composition.

In addition to the γ-cyclodextrin, the ophthalmic composition of thedisclosure may further comprise α-cyclodextrin, β-cyclodextrin and/or awater-soluble cyclodextrin derivative as defined above.

The dexamethasone ophthalmic composition of the disclosure comprises anophthalmically acceptable medium as defined above.

According to a preferred embodiment, the ophthalmically acceptablemedium comprises water and optionally an additive selected from thegroup consisting of a preservative, a stabilizing agent, an electrolyte,a buffering agent, and combinations thereof, as defined above.

In a particularly preferred embodiment, the ophthalmic dexamethasonecomposition comprises:

-   -   1 to 2% of dexamethasone, for example 1.5% of dexamethasone;    -   12 to 16% of γ-cyclodextrin, for example 14% of γ-cyclodextrin;    -   2.2 to 2.8% of polymer, for example 2.5% of poloxamer;    -   0 to 0.2% of stabilizing agent, for example 0.1% of disodium        edetate;    -   0 to 1% of electrolyte, for example 0.57% of sodium chloride;        and    -   water;        wherein the % are % by weight based on the volume of the        composition.

Method of Preparing Ophthalmic Compositions According to the Disclosure

The compositions of the disclosure can be obtainable by or obtained bythe following methods. All of the embodiments, preferred recitations andparticular examples cited in the previous sections equally apply to themethods of the disclosure and the compositions obtained with the methodsof the disclosure.

In a first embodiment, the method of preparing an ophthalmic compositioncomprises the steps of:

-   -   a) suspending an active pharmaceutical ingredient and a        cyclodextrin in an ophthalmically acceptable medium to form a        suspension;    -   b) heating the suspension at a temperature T1 lower than 120° C.        for a time t until the active pharmaceutical ingredient and the        cyclodextrin are substantially dissolved in the ophthalmically        acceptable medium; and    -   c) cooling the resulting solution to a temperature T2 to obtain        an ophthalmic composition comprising a solid complex of an        active pharmaceutical ingredient and a cyclodextrin.

In the method of the first embodiment, the active pharmaceuticalingredient and the cyclodextrin may be suspended in an ophthalmicallyacceptable medium to provide a suspension having a milky appearance. Thesuspension may then be heated for a sufficient time at a sufficienttemperature until both the active pharmaceutical ingredient andcyclodextrin are dissolved in the ophthalmically acceptable medium, andno degradation product is formed. Once the active pharmaceuticalingredient and the cyclodextrin are dissolved, the milky suspension mayturn into a substantially clear solution. The resulting solution maythen be cooled at a rate sufficient to produce a microsuspensioncomprising a solid active pharmaceutical ingredient/cyclodextrincomplex.

In a second embodiment, the method of preparing an ophthalmiccomposition comprises the steps of:

-   -   a) suspending a cyclodextrin in an ophthalmically acceptable        medium to form a suspension;    -   b) heating the suspension until the cyclodextrin is        substantially dissolved in the ophthalmically acceptable medium;    -   c) adding an active pharmaceutical ingredient in solid form in        the solution of step b) at a temperature T1 lower than 120° C.        and heating the mixture at a temperature T1 lower than 120° C.        for a time t until the active pharmaceutical ingredient is        substantially dissolved in the ophthalmically acceptable medium;        and    -   d) cooling the resulting solution to a temperature T2 to obtain        an ophthalmic composition comprising a solid complex of an        active pharmaceutical ingredient and a cyclodextrin.

In the method of the second embodiment, a cyclodextrin may be suspendedin an ophthalmically acceptable medium to provide a suspension having amilky appearance. The cyclodextrin suspension may be heated for asufficient time at a sufficient temperature until the cyclodextrin isdissolved in the ophthalmically acceptable medium. An activepharmaceutical ingredient may be added in solid form to the heatedaqueous solution, while stirring the solution. The heating may becarried out for a sufficient time at a sufficient temperature until theactive pharmaceutical ingredient is dissolved in the ophthalmicallyacceptable medium, and no degradation product is formed. The resultingsolution may be cooled at a rate sufficient to produce a microsuspensioncomprising a solid active pharmaceutical ingredient/cyclodextrincomplex.

In a third embodiment, the method of preparing an ophthalmic compositioncomprises the steps of:

-   -   a) suspending an active pharmaceutical ingredient in an        ophthalmically acceptable medium to form a suspension and        heating said suspension until the active pharmaceutical        ingredient is substantially dissolved in the ophthalmically        acceptable medium;    -   b) suspending a cyclodextrin in an ophthalmically acceptable        medium to form a suspension and heating said suspension until        the cyclodextrin is substantially dissolved in the        ophthalmically acceptable medium;    -   c) mixing the compositions of step a) and b) at a temperature T1        lower than 120° C. and heating the mixture at a temperature T1        lower than 120° C. for a time t; and    -   d) cooling the resulting solution to a temperature T2 to obtain        an ophthalmic composition comprising a solid complex of an        active pharmaceutical ingredient and a cyclodextrin.

In the method of the third embodiment, an active pharmaceuticalingredient may be suspended in an ophthalmically acceptable medium freeof cyclodextrin. The resulting suspension may have a milky appearance.Separately a cyclodextrin may be suspended in an ophthalmicallyacceptable medium free of active pharmaceutical ingredient. Theresulting suspension may have a milky appearance. The two suspensionsmay be heated or sterilized by, for example, heating in an autoclave for121° C. for 20 minutes. Then the two suspensions or hot solutions may bemixed together and the mixture may be heated until the complex of activepharmaceutical ingredient and cyclodextrin is formed, and no degradationproduct is formed. The resulting solution may be cooled at a ratesufficient to produce a microsuspension comprising a solid activepharmaceutical ingredient/cyclodextrin complex.

The ophthalmic composition obtained with the methods of the first,second and third embodiments may comprise less than 2%, in particularless than 1%, more particularly less than 0.8%, by weight of impuritiesbased on the weight of the active pharmaceutical ingredient.

The present disclosure also provides methods for preparing dexamethasoneophthalmic compositions according to the disclosure.

Accordingly, in a fourth embodiment, the method of preparing anophthalmic composition, comprising the steps of:

-   -   a) suspending dexamethasone and γ-cyclodextrin in an        ophthalmically acceptable medium to form a suspension;    -   b) heating the suspension at a temperature T1 lower than 120° C.        for a time t until the dexamethasone and the γ-cyclodextrin are        substantially dissolved in the ophthalmically acceptable medium;        and    -   c) cooling the resulting solution to a temperature T2 to obtain        an ophthalmic composition comprising a solid complex of        dexamethasone and γ-cyclodextrin.

In a fifth embodiment, the method of preparing an ophthalmic compositioncomprises the steps of:

-   -   a) suspending γ-cyclodextrin in an ophthalmically acceptable        medium to form a suspension;    -   b) heating the suspension until the γ-cyclodextrin is dissolved        in the ophthalmically acceptable medium;    -   c) adding dexamethasone in solid form in the solution of step b)        at a temperature T1 lower than 120° C. and heating the mixture        at a temperature T1 lower than 120° C. for a time t until the        dexamethasone is substantially dissolved in the ophthalmically        acceptable medium; and    -   d) cooling the resulting solution to a temperature T2 to obtain        an ophthalmic composition comprising a solid complex of        dexamethasone and γ-cyclodextrin.

In a sixth embodiment, the method of preparing an ophthalmic compositioncomprises the steps of:

-   -   a) suspending dexamethasone in an ophthalmically acceptable        medium to form a suspension and heating said suspension until        the dexamethasone is substantially dissolved in the        ophthalmically acceptable medium;    -   b) suspending γ-cyclodextrin in an ophthalmically acceptable        medium to form a suspension and heating said suspension until        the γ-cyclodextrin is substantially dissolved in the        ophthalmically acceptable medium;    -   c) mixing the compositions of steps a) and b) at a temperature        T1 lower than 120° C. and heating the mixture at a temperature        T1 lower than 120° C. for a time t; and    -   d) cooling the resulting solution to a temperature T2 to obtain        an ophthalmic composition comprising a solid complex of        dexamethasone and γ-cyclodextrin.

The ophthalmic compositions obtained with the methods of the third,fourth and fifth embodiments may comprise less than 0.5%, in particularless than 0.3%, more particularly less than 0.2%, by weight of16,17-unsaturated dexamethasone and dexamethasone enol aldehydes basedon the weight of dexamethasone.

The heating step of the methods of the disclosure is carried out at atemperature T1 lower than 120° C. so as to avoid generation ofimpurities. In particular, temperature T1 may be from 80 to 110° C.,more particularly from 85 to 105° C., even more particularly from 90 to100° C.

The heating step of the methods of the disclosure is carried out for atime t. In particular, the heating time t is from 5 minutes to 2 hours,more particularly from 10 minutes to 1 hour, even more particularly from15 to 30 minutes.

During the heating cycle, the active pharmaceutical ingredient and/orthe cyclodextrin are dissolved and the complex of active pharmaceuticalingredient and cyclodextrin is formed. Heating is accomplished by anymethod or means known to those having ordinary skill in the art. Inpreferred embodiments, the heating is accomplished with an autoclave ora jacketed reactor with steam.

The cooling step of the methods of the disclosure lowers the temperatureof the composition from temperature T1 to temperature T2 in order toprecipitate the solid complex of active pharmaceutical ingredient andcyclodextrin. In particular, temperature T2 may be from 10 to 40° C.,more particularly from 15 to 35° C., even more particularly from 20 to30° C.

The cooling rate of the methods of the disclosure may be carried bylowering temperature T1 to temperature T2 with a rate of 1 to 25°C./min, in particular 2 to 20° C./min, more particularly 5 to 18°C./min.

The cooling may be accomplished by any method or means known to thosehaving ordinary skill in the art. In preferred embodiments, the coolingis accomplished with an ice bath or a jacketed reactor with a coolant.

In the methods of some exemplary embodiments, for example, the first,second, third, fourth, fifth and sixth embodiments, the suspension ofstep a) may further comprise a polymer as defined above. In the methodsof the third and sixth embodiments, the suspension of step b) mayfurther comprise a polymer as defined above. When the initial suspensioncomprises a polymer, part of the polymer may be taken up within thesolid complex and/or part of the polymer may be coated on the surface ofthe solid complex. The microsuspension obtained with the method of thedisclosure may thus comprise a microparticle drug/cyclodextrin/polymercomplex. Said drug/cyclodextrin/polymer complex may comprise a polymercoating.

When a polymer is introduced in the initial suspension of the methods ofthe disclosure, the ophthalmic composition obtained with said methodsmay exhibit a viscosity of 4 to 14 cP, preferably 5 to 13 cP, morepreferably 6 to 12 cP.

The viscosity of the compositions obtained with the methods of thedisclosure is higher than that of compositions obtained with knownmanufacturing methods. Without wishing to be bound by theory, Applicantsbelieve that the implementation of the controlled cooling step of themethod, in particular with a cooling rate of 1 to 25° C./min, after theheating step allows less polymer to be included in the solid complex,and therefore more polymer is found in solution, thereby increasing theviscosity of the formulation. Hence, the original manufacturing processdisclosed in the present application enables to obtain new formulationswith increased viscosity with similar amounts of polymer, cyclodextrinand active pharmaceutical ingredient, as prior art formulations.

In the methods of the exemplary embodiments, for example, the first,second, third, fourth, fifth and sixth embodiments, the ophthalmicallyacceptable medium may comprise water and optionally an additive selectedfrom the group consisting of a preservative, a stabilizing agent, anelectrolyte, a buffering agent, and combinations thereof.

In exemplary methods, for example, the first, second, fourth and fifthembodiments, the ophthalmically acceptable medium of step a) maycomprise water and optionally an additive selected from the groupconsisting of a preservative, a stabilizing agent, an electrolyte, abuffering agent, and combinations thereof.

In further exemplary methods, for example, the third and sixthembodiments, the ophthalmically acceptable medium of step a) maycomprise only water and the ophthalmically acceptable medium of step b)may comprise water and optionally an additive selected from the groupconsisting of a preservative, a stabilizing agent, an electrolyte, abuffering agent, and combinations thereof.

In alternative embodiments, for example, the methods of the third andsixth embodiments, the ophthalmically acceptable medium of step b) maycomprise only water and the ophthalmically acceptable medium of step a)may comprise water and optionally an additive selected from the groupconsisting of a preservative, a stabilizing agent, an electrolyte, abuffering agent, and combinations thereof.

Uses of the Composition of the Disclosure

The ophthalmic compositions of the disclosure may be for use in thetreatment of an ocular condition, in particular an anterior ocularcondition or a posterior ocular condition, more particularly uveitis,macular edema, macular degeneration, retinal detachment, ocular tumors,fungal or viral infections, multifocal choroiditis, diabeticretinopathy, proliferative vitreoretinopathy (PVR), sympatheticophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uvealdiffusion, and vascular occlusion. The compositions of the disclosuremay be particularly useful in treating uveitis, macular edema, diabeticretinopathy, proliferative vitreoretinopathy (PVR), and vascularocclusions.

The dexamethasone ophthalmic composition according to the disclosure mayin particular be used for the treatment of macular edema. In this case,the dexamethasone ophthalmic composition according to the disclosure maybe topically administered to the eye in an amount of 1 drop ofcomposition three times per day. The amount of dexamethasone in saidcomposition may be from 1 to 2%, in particular 1.5% by weight ofdexamethasone based on the volume of the composition.

The compositions of the disclosure do not need to be administered asfrequently as know topical dexamethasone compositions, i.e. 1 drop ofcomposition six times per day. Indeed, due to the enhanced viscosity ofthe composition, the solid complexes of the composition of thedisclosure exhibit higher contact time on the surface of the eyecompared to known compositions which increases the bioavailability ofthe active pharmaceutical ingredient.

The present disclosure also covers the use of the ophthalmic compositionof the disclosure as an eye drop solution.

Measuring Methods Diameter

The diameter of a particle, such as a solid complex of activepharmaceutical ingredient and cyclodextrin, can correspond to the D₅₀diameter of the particle. Diameter D₅₀ is also known as the mediandiameter or the medium value of the particle size distribution. DiameterD₅₀ corresponds to the value of the particle diameter at 50% in thecumulative distribution. For example, if D₅₀ is 5 μm, then 50% of theparticles in the sample are larger than 5 μm, and 50% smaller than 5 μm.Diameter D₅₀ is usually used to represent the particle size of a groupof particles.

The diameter and/or size of a particle or complex can be measuredaccording to any method known to those of ordinary skill in the art. Forexample, the diameter D₅₀ is measured by laser diffraction particle sizeanalysis. Generally, there are a limited number of techniques formeasuring/evaluating cyclodextrin/drug particle or complex diameterand/or size. In particular, persons of ordinary skill in this field knowthat the physical properties (e.g. particle size, diameter, averagediameter, mean particle size, etc.) are typically evaluated/measuredusing such limited, typical known techniques. For example, such knowntechniques are described in Int. J. Pharm. 493(2015), 86-95, cited abovein paragraph [00076], which is incorporated by reference herein in itsentirety. In addition, such limited, known measurement/evaluationtechniques were known in the art as evidenced by other technicalreferences such as, for example, European Pharmacopoeia (2.9.31 Particlesize analysis by laser diffraction, January 2010), and Saurabh Bhatia,Nanoparticles types, classification, characterization, fabricationmethods and drug delivery applications, Chapter 2, Natural Polymer DrugDelivery Systems, PP. 33-94, Springer, 2016, which are also incorporatedby reference herein in their entireties.

For particle size of complexes comprising an active pharmaceuticalingredient other than dexamethasone, the particle size is measured bylaser diffraction particle size analysis according to Pharm. Eur.2.9.31.

For particle size of complexes comprising dexamethasone, the particlesize is measured by laser diffraction particle size analysis accordingto Pharm. Eur. 2.9.31 with the following parameters:

-   -   System: Malvern Mastersizer 3000 with hydro MV disperser    -   Fraunhofer approximation    -   Dispersant: water    -   Refractive index of the dispersant: 1.33    -   Time of measurement: 1 second    -   Time of measurement of the background: 10 second    -   Stirrer speed: 1200 rpm    -   Obscuration range: 1-20%    -   Model: standard    -   Sample preparation: Homogenize the eye drops by shaking    -   Sample size: addition of 0.5 ml eye drop to the disperser    -   Cleaning: rinsing twice with the dispersant (water) and start a        measurement, checking that beam strength is less than 120 units        in the first channels, and loading a background.

Viscosity

The viscosity of a composition corresponds to the dynamic viscosity ofsaid composition. The viscosity is measured at 25° C. with a Brookfielddigital viscometer. The viscosity of a composition is measured shortlyafter, i.e. less than 24 hours after, the preparation of thecomposition.

Percentage of Drug in Solid Complex and Percentage of Dissolved Drug

The amount of drug in the form of solid complexes and the amount ofdissolved drug is obtained by centrifuging the composition at 6000 rpmat a temperature of 22-230 C for 20-30 minutes.

The amount of dissolved drug corresponds to the amount of drug in thesupernatant as measured by high-performance liquid chromatography. Thepercentage of drug in the form of a solid complex is obtained with thefollowing formula:

${\%{{drug}{in}{solid}{complex}}} = {\frac{\left( {{{total}{drug}} - {{dissolved}{drug}}} \right)}{{total}{drug}} \times 100}$

wherein“total drug” is the total amount of drug introduced in the compositionin mg/mL; and “dissolved drug” is the amount of drug in the supernatantin mg/mL.

The percentage of dissolved drug is obtained with the following formula:

% dissolved drug=100−% drug in solid complex

EXAMPLES

The following Examples are detailed by way of illustration only and arenot to be construed as limiting in spirit or in scope, manymodifications both in materials and in methods will be apparent to thoseskilled in the art.

Example 1

The composition of the aqueous dexamethasone eye drops are as follows:dexamethasone (1.50%), γ-cyclodextrin (14.00%), poloxamer 407 (2.50%),benzalkonium chloride (0.02%), disodium edetate (0.10%), sodium chloride(0.57%) in purified water, all w/v %. Five different methods areapplied.

F1: dissolving or suspending the ingredients, including dexamethasone,in pure water and autoclaving the mixture in sealed vial at 121° C. for20 minutes. The vials comprising a substantially clear aqueous solutionare removed from the autoclave, and become cloudy with cooling toambient temperature. The solid particles are analyzed by Fouriertransform infra-red spectroscopy (FTIR), differential scanningcalorimetry (DSC) and x-ray diffraction (XRD) indicating that the solidparticles comprise dexamethasone/γ-cyclodextrin complexes.

F2: dissolving or suspending the ingredients, including dexamethasone,in pure water and heating the mixture over 30 minutes to 90° C. to forma clear solution. Further heating at 90° C. for 15 minutes the solutionis allowed cool and become cloudy at ambient temperature reaching roomtemperature within approximately 3 hours.

F3: dissolving or suspending the pharmaceutical excipients in purewater, heating the mixture to 90° C. to form clear solution and thenadding solid dexamethasone powder to the hot solution. Whendexamethasone is dissolved (under stirring for 15 minutes) the solutionis allowed cool and become cloudy at ambient temperature reaching roomtemperature within approximately 3 hours.

F4: dissolving or suspending the pharmaceutical excipients in pure waterand autoclaving the mixture in sealed vial at 121° C. for 20 minutes toform clear solution. After cooling to 95° C. solid dexamethasone powderis added to the solution. When dexamethasone is dissolved (understirring for 15 minutes) the solution is allowed cool and become cloudyat ambient temperature reaching room temperature over approximately 3hours.

F5: dissolving or suspending the pharmaceutical excipients in pure waterand autoclaving the mixture in sealed vial at 121° C. for 20 minutes toform clear solution. After cooling to 95° C. solid dexamethasone powderis added to the solution. When dexamethasone is dissolved (understirring for 15 minutes) the solution was rapidly cooled to roomtemperature (over 20 minutes) and becomes cloudy with cooling.

F6: the excipients were separated into two parts, A and B. In part A,all the excipients except γ-cyclodextrin were dissolved in pure water at80° C. and, in part B γ-cyclodextrin was suspended (or dissolved)separately in pure water at 80° C. The dexamethasone was added to theexcipient mixture just before sterilization. The two parts of theexcipient mixture in water containing dexamethasone (part A) andγ-cyclodextrin suspended (or dissolved) in water (part B), weresterilized at 121° C. for 15 minutes. After sterilization, the sterileγ-cyclodextrin was added to the rest of the sterile excipients at 95° C.In other words, parts A and B were mixed. After stirring for 15 minutesthe solution was rapidly cooled to room temperature (over 20 minutes) toform cloudy suspension. F6 did not contain benzalkonium chloride.

TABLE 1 Results of the microparticle formation studies. Mean of threedeterminations ± standard deviation. Formulation F1 F2 F3 F4 F5 F6 pH4.5 5.1 5.0 5.0 4.7 4.7 Solid dexamethasone 87.6 ± 0.0  70.8 ± 0.0  68.1± 0.1  73.0 ± 0.2  87.1 ± 0.1  83.9 ± 0.6  fraction (%) Solidγ-cyclodextrin 91.6 ± 0.0  89.6 ± 3.4  90.1 ± 3.4  88.8 ± 0.0  89.9 ±0.0  — fraction (%) Viscosity at 25° C. (cP) 3.68 ± 0.29 6.48 ± 0.076.64 ± 0.12 8.18 ± 0.05 8.67 ± 0.30 11.1 ± 0.1  Mean particle size(μm)4.4 ± 2.7 3.6 3.3

The results show that rapid cooling (F5) gives microsuspension wheremore than about 80% of the active pharmaceutical ingredient andγ-cyclodextrin are in the solid phase as active pharmaceuticalingredient/γ-cyclodextrin complexes and where most of the polymer is inthe aqueous solution (i.e. have the highest viscosity). F1 settles overtime, but is capable of being re-dispersed with some agitation. However,F5 has a low tendency to settle over time and is readily re-dispersedwith agitation. Thus, F5 displays significant greater physical stabilitythan F1.

Example 2

The composition of the aqueous irbesartan eye drops is as follows:irbesartan (2.0%), γ-cyclodextrin (10.0%), HPMC (0.20%), tyloxapol(0.10%), benzalkonium chloride (0.02%), disodium edetate (0.10%), sodiumchloride (0.50%) in purified water, all w/v %. Three different methodsare applied:

F7: dissolving or suspending the ingredients, including irbesartan, inpure water and autoclaving the mixture in sealed vial at 121° C. for 20minutes to form clear solution. The clear aqueous solution is allowed tocool to ambient temperature and become cloudy.

F8: dissolving or suspending the pharmaceutical excipients in pure waterand autoclaving the mixture in sealed vial at 121° C. for 20 minutes toform clear solution. After cooling to 95° C. solid irbesartan powder isadded to the solution. When irbesartan was dissolved (15 minutes) thesolution is allowed cool and become cloudy at ambient temperaturereaching room temperature within approximately 3 hours.

F9: dissolving or suspending the pharmaceutical excipients in pure waterand autoclaving the mixture in sealed vial at 121° C. for 20 minutes toform clear solution. After cooling to 95° C. solid irbesartan powder isadded to the solution. When irbesartan is dissolved (15 minutes) thesolution is rapidly cooled to room temperature (within 20 minutes) andbecame cloudy.

For formulation F7, the formulation can have a solid drug fraction of54%, a viscosity at 25° C. (cP) of 4.36 and a mean particle size (μm) of2.44.

Example 3

The dehydration of dexamethasone during preparation of the aqueousdexamethasone eye drops described in Example 1 (formulations F1, F2, F3,F4, F5 and F6) is determined by quantitative determination of16,17-unsaturated dexamethasone and the dexamethasone enol aldehydes inthe eye drops after manufacturing. Formulations F12, F13 and F14 areprepared as described in F1, that is by dissolving or suspending theingredients, including dexamethasone, in pure water and autoclaving themixture in sealed vial at 121° C. for 20 minutes. The substantiallyclear aqueous vials are removed from the autoclave and become cloudyupon cooling at ambient conditions. The composition of F12 is identicalto F1, but does not contain γ-cyclodextrin. F13 comprises dexamethasoneand γ-cyclodextrin suspended in pure water. F14 comprises dexamethasonesuspended in pure water. The effect of excipients and preparationmethods on the formation of 16,17-unsaturated dexamethasone and thedexamethasone enol aldehydes is presented as the fraction (in %) ofdexamethasone degraded to form 16,17-unsaturated dexamethasone and thedexamethasone enol aldehydes.

TABLE 2 Formulation F1 F2 F3 F4 F5 F6 F12 F13 F14 Autoclaving at 121° C.X X X X X for 20 min. with dexamethasone Heating to 90° C. for X X X X X15 min. with dexamethasone Comprises Yes Yes Yes Yes Yes Yes No Yes Noγ-cyclodextrin pH 4.5 5.1 5.0 5.0 4.7 4.8 Fraction of 12.4 29.2 31.927.0 12.9 16.1 1.17 4.16 5.73 dexamethasone in solution (%) Fractiondegraded 1.45 0.16 0.11 0.11 0.08 0.20 0.18 1.34 0.13 to16,17-unsaturated dexamethasone and enol aldehydes (%)

The results presented in Table 2 show that formation of16,17-unsaturated dexamethasone and dexamethasone enol aldehydes iscatalyzed by the presence of γ-cyclodextrin during heating in anautoclave, formulation F1 (comprises all the excipients) and F13(comprises γ-cyclodextrin but not the other excipients). Much less16,17-unsaturated dexamethasone and the dexamethasone enol aldehydes areformed in the aqueous eye drop formulation when γ-cyclodextrin isremoved from the formulation (F12) or when the eye drops are prepared byheating to 90° C. for 15 minutes (F2, F3 and F4). Little or no16,17-unsaturated dexamethasone and dexamethasone enol aldehydes areformed in the eye drops during storage at room temperature (22-23° C.)for over 12 months. During formulation of F6 aqueous suspension ofγ-cyclodextrin and separately aqueous suspension containingdexamethasone and all other ingredients except γ-cyclodextrin wereautoclaved at 121° C. for 15 minutes. After autoclaving the twosuspensions/solutions were cooled to 95° C. before mixing and thencooled further to ambient temperature (see Example 1). Only minor amountof 16,17-unsaturated dexamethasone and the dexamethasone enol aldehydesis formed in the aqueous eye drop formulation when dexamethasone issterilized in absence of γ-cyclodextrin.

Example 4

The amount of 16,17-unsaturated dexamethasone and dexamethasone enolaldehydes formed, presented as the fraction (in %) of dexamethasonedegraded to form 16,17-unsaturated dexamethasone and dexamethasone enolaldehydes, in aqueous eye drops after no autoclaving (i.e., heating to121° C. for 20 minutes) once, twice and three times is shown in Table 3.

TABLE 3 Number of autoclaving cycles pH 0 1 2 3 2.5 0.08% 1.06% 2.29%3.12% 4.0 0.08% 1.14% 2.19% 3.03% 5.5 0.08% 1.22% 2.27% 3.46% 7.0 0.08%0.86% 1.64% 2.57%

The amount of 16,17-unsaturated dexamethasone and dexamethasone enolaldehydes formed, presented as the fraction (in %) of dexamethasonedegraded to form 16,17-unsaturated dexamethasone and dexamethasone enolaldehydes, in the aqueous eye drops after no autoclaving (i.e. heatingto 121° C. for 20 minutes), once, twice and three times is shown below.

Formation of 16,17-unsaturated dexamethasone and dexamethasone enolaldehydes is observed at all four pH tested. Although, less16,17-unsaturated dexamethasone and dexamethasone enol aldehydes areformed at pH 7.0 than at 5.5 other degradation products appeared.According to the British Pharmacopoeia 2015 (version 19.0) the pH ofaqueous dexamethasone eye drop suspension should be between 5.0 and 6.0.

Example 5

The degradation of dexamethasone in F1 was investigated at 25° C., 40°C., 60° C., 70° C. and 95° C. The apparent first-order rate constantsfor the dexamethasone disappearance were determined, and the apparentactivation energy calculated with the help of Arrhenius Equation. Theequation was also used to estimate the first-order rate constant for thedexamethasone disappearance at 25° C. The time for 10% (t₉₀; theshelf-life) and 0.5% (t_(99.5)) degradation was also calculated from therate constants.

TABLE 4 The first-order rate constant for the degradation ofdexamethasone in aqueous eye drop formulation comprising γ-cyclodextrin(i.e. F5) at pH 7.0. 25° C. 40° C. 60° C. 70° C. 95° C. First-order rate1.442 · 10⁻⁷ 2.483 · 10⁻⁶ 2.476 · 10⁻⁵ 2.044 · 10⁻⁴ 3.277 · 10⁻³constant (k in h⁻¹) Shelf-life 30,444 1,768 177 21.5 1.3 (t₉₀ in days)Time for 0.5% 34,761 2,018 202 24.5 1.5 degradation (t_(99.5) in hours)Apparent activation energy (E_(a))${32.2\frac{kcal}{mol}} = {134.5\frac{kJ}{mol}}$ Estimated shelf- 83.4years life (t₉₀) at 25° C. Estimated time for  4.0 years 0.5%degradation (t_(99.5)) at 25° C.

In general, values of E_(a) range from about 50 to 85 kJ/mol and valuesgreater than 100 kJ/mol are very uncommon. The apparent value of E_(a)for dehydration of dexamethasone in aqueous γ-cyclodextrin solution toform 16,17-unsaturated dexamethasone and the dexamethasone enolaldehydes is 134.5 kJ/mol and, thus, 16,17-unsaturated dexamethasone andthe dexamethasone enol aldehydes are only formed in aqueousγ-cyclodextrin solutions at exceptionally high temperatures.

Example 6

The effect of the cooling rate on the size of the microparticles isexamined. The dexamethasone eye drop vehicle contained containsγ-cyclodextrin (14.00%), poloxamer 407 (2.50%), benzalkonium chloride(0.02%), disodium edetate (0.10%), sodium chloride (0.57%) in purifiedwater, all w/v %. The vehicle is heated in an autoclave (121° C. for 20min) in sealed vial to form a substantially clear solution. Aftercooling to 95° C. solid dexamethasone powder (1.50% w/v) is added to thesolution. After dexamethasone is dissolved (under stirring for 15minutes), the solution is divided into small portions (approx. 5 ml) andis placed into thermostated water set to different temperatures. Thetemperature changes are recorded against time. The particle size and theviscosity of the resulting suspensions are determined.

Example 7

The effect of sterilization time and temperature, and the effect ofmixing time, is examined in the case of formulation F6. Formulations areprepared as described in Example 1. During formulation of F6 aqueoussuspension of γ-cyclodextrin (part B) and separately aqueous suspensioncontaining dexamethasone and all other ingredients except γ-cyclodextrin(part A) are autoclaved at 121° C. for 15 minutes. After autoclaving thetwo suspensions/solutions are cooled to 95° C. before mixing and thenare cooled further to ambient temperature (see Example 1 and Example 3).

The modified technological parameters during sterilization and mixing ofγ-cyclodextrin to the other excipients and the resulting amounts offraction degraded to 16,17-unsaturated dexamethasone and dexamethasoneenol aldehydes are shown in Table 5.

TABLE 5 Amount of 16,17- unsaturated Mixing dexamethasone with and theγCD dexamethasone soln. enol Formulation Sterilization at 95° C.aldehydes (%) F6 15 min, 121° C. 15 min 0.20 F6a 15 min, 121° C. No γCD0.05 F6b 2 × 15 min, 121° C. No γCD 0.10 F6c 15 min, 121° C. 30 min 0.28F6d 2 × 15 min, 121° C. 15 min 0.25 F6e 15 min, 135° C. 15 min 0.34

Formulation F6a and F6b do not contain any γCD and show only the effectof the sterilization step. F6a was sterilized in an autoclave once for15 min at 121° C. while F6b went through two sterilization cycles at thesame conditions. One sterilization cycle added 0.05% of16,17-unsaturated dexamethasone and dexamethasone enol aldehydes (% ofthe total amount of dexamethasone in the eye drops) and twosterilization cycles added 0.10% of 16,17-unsaturated dexamethasone anddexamethasone enol aldehydes.

Formulation F6c shows the effect of double mixing time at 95° C. afterthe addition of γCD. An extra 15 minute of mixing adds an extra 0.08% of16,17-unsaturated dexamethasone and dexamethsone enol aldehydes to the0.20% of F6.

Formulation F6d shows the effect of double autoclaving (twosterilization cycles) combined together with the 15 minutes mixing at95° C. after the addition of γCD. The double sterilization added anextra 0.05% of 16,17-unsaturated dexamethasone and dexamethasone enolaldehydes to the 0.20% of F6, which corresponds with the results of theF6a and F6b formulations not containing γCD.

Formulation F6e shows the effect of sterilizing at 135° C. instead of121° C. for 15 minutes, and then 15 min mixing at 95° C. after theaddition of γCD. The amount of 16,17-unsaturated dexamethasone anddexamethasone enol aldehydes increased from 0.20% to 0.34%.

The results show that the manufacturing method is robust and smallchanges in the technological parameters will essentially not affect theamount of 16,17-unsaturated dexamethasone and dexamethasone enolaldehydes in the final product.

Example 8

Manufacturing process F6 was implemented on industrial scale. Thecomposition of the aqueous dexamethasone eye drops are as follows:dexamethasone (1.50% w/v), γ-cyclodextrin (14.00% w/v), poloxamer 407(2.50% w/v), disodium edetate (0.10% w/v), sodium chloride (0.57% w/v)in purified water The batch size was 400 liters (F15).

F15: Dexamethasone and all the excipients except γ-cyclodextrin weredissolved or suspended in pure water at 80° C. (Solution A).γ-Cyclodextrin was dissolved separately in pure water at 80° C.(Solution B). Solution A and Solution B were sterilized at 121° C. for15 minutes. After sterilization and cooling Solution A and Solution Bwere mixed. The mixture was then reheated to 95° C. and kept at thattemperature under stirring for 15 minutes. The solution was cooled to40° C. in under 40 minutes and then to room temperature (within another40 minutes). Then the resulting microsuspension upon was filled intounit dose containers. See Table 6.

TABLE 6 Formulation Parameter F15 pH 4.5 D₅₀ (μm) 7.2 Fraction degradedto 16,17- 0.3% unsaturated dexamethasone and dexamethasone enolaldehydes Uniformity of dosage units (L1 < 15 L1 = 3.9 according toPh.Eur.)

Example 9

The composition of the aqueous dexamethasone eye drops (F16) are asfollows: dexamethasone (1.50%), γ-cyclodextrin (14.00%), poloxamer 407(2.50%), disodium edetate (0.10%), sodium chloride (0.57%) in purifiedwater, all w/v %. The excipients were separated into two parts, A and B.In A all the excipients except γ-cyclodextrin were dissolved in purewater at 80° C. and in B γ-cyclodextrin was dissolved separately in purewater at 80° C. The dexamethasone was added to the excipient mixturejust before sterilization. The excipient mixture in water containingdexamethasone (A) and γ-cyclodextrin dissolved in water (B), weresterilized at 121° C. for 15 minutes. After sterilization, A and B weremixed at 95° C. After stirring for 15 minutes the solution was cooledfrom 95° C. to 40° C. at three different cooling rates (ΔT/Δt). Thecooling rate of F16a was 17.7° C./min, that of F16b 1.3° C./min and thatof F16c 1.2° C./min (Table 7).

TABLE 7 F16a F16b F16c ΔT/Δt (° C./min) 17.7 1.3 1.2 D₅₀ (μm) 3.3 7.28.8 Viscosity (cP) 11.1 9.0 not determined

The table shows how the mean particle size is controlled by the coolingrate. The faster the cooling rate, the smaller the particles and thehigher the viscosity.

Example 10

Kinase inhibitors were suspended in aqueous solutions containing from 1%(w/v) to 15% (w/v) γ-cyclodextrin. Heating of the suspensions formed inan autoclave (121° C. for 15 min) resulted in up to 50% degradation ofthe kinase inhibitors whereas heating to 95° C. for 15 min and rapidcooling to 25° C. resulted in significant less degradation and producedmicroparticle suspension. The solubility of the kinase inhibitors inpure water (S₀) and the stability constant (K_(1:1)) of the activepharmaceutical ingredient/γ-cyclodextrin complexes were determined fromthe initial linear art of the phase-solubility profiles (Table 8).

TABLE 8 Kinase Inhibitor S₀ (mg/mL) K_(1:1) (M⁻¹) Axitinib 0.0004 260Cediranib 1.2 23 Dovitinib 0.006 680 Motesanib 0.014 140 Pazopanib0.0006 13 Regorafenib 0.0001 94

Example 11

Kinase inhibitors were suspended in aqueous media consisting of purewater, aqueous γ-cyclodextrin solution, aqueous solution containingγ-cyclodextrin and tyloxapol (1% w/v), and aqueous eye drop mediumcontaining γ-cyclodextrin, benzalkonium chloride (0.02% w/v), disodiumedetate (0.10% w/v) and sodium chloride (0.05% w/v). The concentrationof γ-cyclodextrin varied depending on the kinase inhibitor. Thesolubility was determined as described in Example 10. Table 9 shows theγ-cyclodextrin concentration and the effect of tyloxapol and the eyedrop excipient mixture on the γ-cyclodextrin solubilization of thekinase inhibitors.

TABLE 9 γ-Cyclodextrin Solubility (mg/mL) Kinase concentration γ- EyeInhibitor (% w/v) Cyclodextrin Tyloxapol drops Axitinib 4 0.003 0.0080.007 Cediranib 12 2.0 2.8 4.1 Motesanib 8 0.09 0.04 0.07 Pazopanib 160.007 0.006 0.01 Regorafenib 4 0.0005 0.005 0.0008

The results show that the excipients can have significant effect on thekinase inhibitor solubilization.

Example 12

Dovitinib is suspended in aqueous solutions containing from 1% (w/v) to15% (w/v) γ-cyclodextrin. Heating of the suspensions formed in anautoclave (121° C. for 15 min) should result in degradation of dovitinibwhereas heating to 95° C. for 15 min and cooling to 25° C. over 20minutes should result in a composition comprising soliddovitinib/γ-cyclodextrin complexes and significantly less degradation ofdovitinib.

Example 13

Losartan is suspended in aqueous solutions containing from 1% (w/v) to15% (w/v) γ-cyclodextrin. Heating of the suspensions formed in anautoclave (121° C. for 15 min) should result in degradation of losartanwhereas heating to 95° C. for 15 min and cooling to 25° C. over 20minutes should result in a composition comprising solidlosartan/γ-cyclodextrin complexes and significantly less degradation oflosartan.

Example 14

Olmesartan is suspended in aqueous solutions containing from 1% (w/v) to15% (w/v) γ-cyclodextrin. Heating of the suspensions formed in anautoclave (121° C. for 15 min) should result in degradation ofolmesartan whereas heating to 95° C. for 15 min and cooling to 25° C.over 20 minutes should result in a composition comprising solidolmesartan/γ-cyclodextrin complexes and significantly less degradationof olmesartan.

Example 15

Dorzolamide is suspended in aqueous solutions containing from 1% (w/v)to 15% (w/v) γ-cyclodextrin. Heating of the suspensions formed in anautoclave (121° C. for 15 min) should result in degradation ofdorzolamide whereas heating to 95° C. for 15 min and cooling to 25° C.over 20 minutes should result in a composition comprising soliddorzolamide/γ-cyclodextrin complexes and significantly less degradationof dorzolamide.

Example 16

Diclofenac is suspended in aqueous solutions containing from 1% (w/v) to15% (w/v) γ-cyclodextrin. Heating of the suspensions formed in anautoclave (121° C. for 15 min) should result in degradation ofdiclofenac whereas heating to 95° C. for 15 min and cooling to 25° C.over 20 minutes should result in a composition comprising soliddiclofenac/γ-cyclodextrin complexes and significantly less degradationof diclofenac.

Example 17

Nepafenac is suspended in aqueous solutions containing from 1% (w/v) to15% (w/v) γ-cyclodextrin. Heating of the suspensions formed in anautoclave (121° C. for 15 min) should result in degradation of nepafenacwhereas heating to 95° C. for 15 min and cooling to 25° C. over 20minutes should result in a composition comprising solidnepafenac/γ-cyclodextrin complexes and significantly less degradation ofnepafenac.

Any numbers expressing quantities of ingredients, constituents, reactionconditions, and so forth used in the specification are to be understoodas being modified in all instances by the term “about.” Notwithstandingthat the numerical ranges and parameters setting forth, the broad scopeof the subject matter presented herein are approximations, the numericalvalue set forth are indicated as precisely as possible. Any numericalvalue, however, may inherently contain certain error or inaccuracies asevident from the standard deviation found in their respectivemeasurement techniques. None of the features recited herein should beinterpreted as invoking 35 U.S.C. § 112(f), or pre-AIA ¶6, unless theterm “means” is explicitly used.

Although the present invention has been described in connection withexemplary embodiments thereof, it will be appreciated by those skilledin the art that additions, deletions, modifications, and substitutionsnot specifically described may be made without departing from the spiritand scope of the disclosure.

1.-109. (canceled)
 110. A sterile ophthalmic microsuspension comprising,in an ophthalmically acceptable medium: a solid complex comprisingdexamethasone and γ-cyclodextrin, wherein said solid complex has adiameter D50 from about 1 μm to about 25 μm; and wherein themicrosuspension comprises less than 1% by weight of degradation productsor degradation products, based on weight of the dexamethasone.
 111. Thesterile ophthalmic microsuspension according to claim 110, wherein theamount of γ-cyclodextrin is from 5% to 20% by weight based on the volumeof the microsuspension.
 112. The sterile ophthalmic microsuspensionaccording to claim 110, wherein the amount of γ-cyclodextrin is from 12%to 16% by weight based on the volume of the microsuspension.
 113. Thesterile ophthalmic microsuspension according to claim 110, wherein themicrosuspension comprises less than 0.5% by weight of 16,17-unsaturateddexamethasone or a mixture of dexamethasone enol aldehydes based on theweight of the dexamethasone.
 114. The sterile ophthalmic microsuspensionaccording to claim 110, wherein the microsuspension comprises less than0.5% by weight of dexamethasone enol aldehydes based on the weight ofdexamethasone.
 115. The sterile ophthalmic microsuspension according toclaim 110, wherein the dexamethasone is present in the composition at aconcentration of about 10 mg/mL to about 20 mg/mL.
 116. The sterileophthalmic microsuspension according to claim 110, wherein 60 to 95% byweight of the dexamethasone in the microsuspension is in the form of asolid complex.
 117. The sterile ophthalmic microsuspension according toclaim 110, wherein the microsuspension further comprises a polymer. 118.The sterile ophthalmic microsuspension according to claim 110, whereinthe microsuspension further comprises a polymer and the amount ofpolymer is 1 to 4%, by weight of polymer based on the volume of themicrosuspension.
 119. The sterile ophthalmic microsuspension accordingto claim 117, wherein the polymer is poloxamer.
 120. The sterileophthalmic microsuspension according to claim 110, wherein themicrosuspension comprises: 1% to 2% of dexamethasone; 12% to 16% ofγ-cyclodextrin; 2.2% to 2.8% of a polymer; 0% to 0.2% of a stabilizingagent; 0% to 1% of an electrolyte; and water; wherein the % are % byweight based on the weight of the microsuspension.
 121. The sterileophthalmic composition according to claim 110, wherein themicrosuspension comprises: about 1.5% of dexamethasone; about 14% ofγ-cyclodextrin; about 2.5% of a poloxamer; about 0.1% of disodiumedetate; about 0.57% of sodium chloride; and water; wherein the % are %by weight based on the weight of the composition.
 122. The sterileophthalmic composition according to claim 1, wherein the microsuspensionhas a pH between about 5.0 and about 6.0.